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M3i
Insect Models of Innate Immunity
Publications
2024
Y Fu, F Jiang, X Zhang, Y Pan, R Xu, X Liang, X Wu, X Li, K Lin, R Shi, X Zhang, D Ferrandon, J Liu, D Pei, J Wang, T Wang
Perturbation of METTL1-mediated tRNA N7-methylguanosine modification induces senescence and aging Journal Article
In: Nature Communications, vol. 8, iss. 15, no. 1, pp. 5713, 2024.
Abstract | Links | BibTeX | Tags: ferrandon, Gene Expression, M3i, RNA, Senescence
@article{Y2024,
title = {Perturbation of METTL1-mediated tRNA N7-methylguanosine modification induces senescence and aging},
author = {Fu Y AND Jiang F and Zhang X and Pan Y and Xu R AND Liang X AND Wu X AND Li X AND Lin K AND Shi R AND Zhang X AND Ferrandon D AND Liu J AND Pei D AND Wang J AND Wang T},
doi = {10.1038/s41467-024-49796-8},
year = {2024},
date = {2024-07-08},
journal = {Nature Communications},
volume = {8},
number = {1},
issue = {15},
pages = {5713},
abstract = {Cellular senescence is characterized by a decrease in protein synthesis, although the underlying processes are mostly unclear. Chemical modifications to transfer RNAs (tRNAs) frequently influence tRNA activity, which is crucial for translation. We describe how tRNA N7-methylguanosine (m7G46) methylation, catalyzed by METTL1-WDR4, regulates translation and influences senescence phenotypes. Mettl1/Wdr4 and m7G gradually diminish with senescence and aging. A decrease in METTL1 causes a reduction in tRNAs, especially those with the m7G modification, via the rapid tRNA degradation (RTD) pathway. The decreases cause ribosomes to stall at certain codons, impeding the translation of mRNA that is essential in pathways such as Wnt signaling and ribosome biogenesis. Furthermore, chronic ribosome stalling stimulates the ribotoxic and integrative stress responses, which induce senescence-associated secretory phenotype. Moreover, restoring eEF1A protein mitigates senescence phenotypes caused by METTL1 deficiency by reducing RTD. Our findings demonstrate that tRNA m7G modification is essential for preventing premature senescence and aging by enabling efficient mRNA translation.},
keywords = {ferrandon, Gene Expression, M3i, RNA, Senescence},
pubstate = {published},
tppubtype = {article}
}
Cellular senescence is characterized by a decrease in protein synthesis, although the underlying processes are mostly unclear. Chemical modifications to transfer RNAs (tRNAs) frequently influence tRNA activity, which is crucial for translation. We describe how tRNA N7-methylguanosine (m7G46) methylation, catalyzed by METTL1-WDR4, regulates translation and influences senescence phenotypes. Mettl1/Wdr4 and m7G gradually diminish with senescence and aging. A decrease in METTL1 causes a reduction in tRNAs, especially those with the m7G modification, via the rapid tRNA degradation (RTD) pathway. The decreases cause ribosomes to stall at certain codons, impeding the translation of mRNA that is essential in pathways such as Wnt signaling and ribosome biogenesis. Furthermore, chronic ribosome stalling stimulates the ribotoxic and integrative stress responses, which induce senescence-associated secretory phenotype. Moreover, restoring eEF1A protein mitigates senescence phenotypes caused by METTL1 deficiency by reducing RTD. Our findings demonstrate that tRNA m7G modification is essential for preventing premature senescence and aging by enabling efficient mRNA translation.
S Ji, JA Hoffmann
Toll-9 prevents the proliferation of injected oncogenic cells in adult flies Journal Article
In: Journal of Genetics and Genomics, pp. 1673-8527, 2024.
Links | BibTeX | Tags: Drosophila, hoffmann, M3i, oncogenic cells, Toll-9
@article{JA2024,
title = {Toll-9 prevents the proliferation of injected oncogenic cells in adult flies},
author = {Ji S AND Hoffmann JA},
doi = {doi: 10.1016/j.jgg.2024.07.002},
year = {2024},
date = {2024-07-05},
urldate = {2024-07-05},
journal = {Journal of Genetics and Genomics},
pages = {1673-8527},
keywords = {Drosophila, hoffmann, M3i, oncogenic cells, Toll-9},
pubstate = {published},
tppubtype = {article}
}
J Chen, G Lin, K Ma, Z Li, S Liégeois, D Ferrandon
In: PLoS Pathog., 2024.
Abstract | Links | BibTeX | Tags: Drosophila melanogaster, ferrandon, Innate immune response, M3i, Pseudomonas aeruginosa
@article{J2024,
title = {A specific innate immune response silences the virulence of Pseudomonas aeruginosa in a latent infection model in the Drosophila melanogaster host},
author = {Chen J and Lin G and Ma K and Li Z and Liégeois S and Ferrandon D},
editor = {. . PLoS Pathog. 2024 Jun 4;20(6):e1012252. . eCollection 2024 Ju},
doi = {doi: 10.1371/journal.ppat.1012252},
year = {2024},
date = {2024-06-04},
urldate = {2024-06-04},
journal = {PLoS Pathog.},
abstract = {Microbial pathogenicity often depends on the route of infection. For instance, P. aeruginosa or S. marcescens cause acute systemic infections when low numbers of bacteria are injected into D. melanogaster flies whereas flies succumb much slower to the continuous ingestion of these pathogens, even though both manage to escape from the gut compartment and reach the hemocoel. Here, we have developed a latent P. aeruginosa infection model by feeding flies on the bacteria for a short period. The bacteria stably colonize internal tissues yet hardly cause any damage since latently-infected flies live almost as long as noninfected control flies. The apparently dormant bacteria display particular characteristics in terms of bacterial colony morphology, composition of the outer cell wall, and motility. The virulence of these bacteria can however be reactivated upon wounding the host. We show that melanization but not the cellular or the systemic humoral response is the predominant host defense that establishes latency and may coerce the bacteria to a dormant state. In addition, the lasting activation of the melanization responses in latently-infected flies provides a degree of protection to the host against a secondary fungal infection. Latent infection by an ingested pathogen protects against a variety of homologous or heterologous systemic secondary infectious challenges, a situation previously described for the endosymbiotic Wolbachia bacteria, a guard against viral infections.},
keywords = {Drosophila melanogaster, ferrandon, Innate immune response, M3i, Pseudomonas aeruginosa},
pubstate = {published},
tppubtype = {article}
}
Microbial pathogenicity often depends on the route of infection. For instance, P. aeruginosa or S. marcescens cause acute systemic infections when low numbers of bacteria are injected into D. melanogaster flies whereas flies succumb much slower to the continuous ingestion of these pathogens, even though both manage to escape from the gut compartment and reach the hemocoel. Here, we have developed a latent P. aeruginosa infection model by feeding flies on the bacteria for a short period. The bacteria stably colonize internal tissues yet hardly cause any damage since latently-infected flies live almost as long as noninfected control flies. The apparently dormant bacteria display particular characteristics in terms of bacterial colony morphology, composition of the outer cell wall, and motility. The virulence of these bacteria can however be reactivated upon wounding the host. We show that melanization but not the cellular or the systemic humoral response is the predominant host defense that establishes latency and may coerce the bacteria to a dormant state. In addition, the lasting activation of the melanization responses in latently-infected flies provides a degree of protection to the host against a secondary fungal infection. Latent infection by an ingested pathogen protects against a variety of homologous or heterologous systemic secondary infectious challenges, a situation previously described for the endosymbiotic Wolbachia bacteria, a guard against viral infections.
PJ Thiele, R Mela-Lopez, SA Blandin, D Klug
Let it glow: genetically encoded fluorescent reporters in Plasmodium Journal Article
In: Malaria Journal, vol. 23, no. 114, 2024.
Abstract | Links | BibTeX | Tags: blandin, fluorescent reporters, M3i, Plasmodium
@article{PJ2024,
title = {Let it glow: genetically encoded fluorescent reporters in Plasmodium},
author = {Thiele PJ and Mela-Lopez R and Blandin SA and Klug D},
editor = {Springer Nature},
doi = {10.1186/s12936-024-04936-9},
year = {2024},
date = {2024-04-20},
urldate = {2024-04-20},
journal = {Malaria Journal},
volume = {23},
number = {114},
abstract = {The use of fluorescent proteins (FPs) in Plasmodium parasites has been key to understand the biology of this obligate intracellular protozoon. FPs like the green fluorescent protein (GFP) enabled to explore protein localization, promoter activity as well as dynamic processes like protein export and endocytosis. Furthermore, FP biosensors have provided detailed information on physiological parameters at the subcellular level, and fluorescent reporter lines greatly extended the malariology toolbox. Still, in order to achieve optimal results, it is crucial to know exactly the properties of the FP of choice and the genetic scenario in which it will be used. This review highlights advantages and disadvantages of available landing sites and promoters that have been successfully applied for the ectopic expression of FPs in Plasmodium berghei and Plasmodium falciparum. Furthermore, the properties of newly developed FPs beyond DsRed and EGFP, in the visualization of cells and cellular structures as well as in the sensing of small molecules are discussed.},
keywords = {blandin, fluorescent reporters, M3i, Plasmodium},
pubstate = {published},
tppubtype = {article}
}
The use of fluorescent proteins (FPs) in Plasmodium parasites has been key to understand the biology of this obligate intracellular protozoon. FPs like the green fluorescent protein (GFP) enabled to explore protein localization, promoter activity as well as dynamic processes like protein export and endocytosis. Furthermore, FP biosensors have provided detailed information on physiological parameters at the subcellular level, and fluorescent reporter lines greatly extended the malariology toolbox. Still, in order to achieve optimal results, it is crucial to know exactly the properties of the FP of choice and the genetic scenario in which it will be used. This review highlights advantages and disadvantages of available landing sites and promoters that have been successfully applied for the ectopic expression of FPs in Plasmodium berghei and Plasmodium falciparum. Furthermore, the properties of newly developed FPs beyond DsRed and EGFP, in the visualization of cells and cellular structures as well as in the sensing of small molecules are discussed.
I Iacobucci, V Monaco, A Hovasse, B Dupouy, R Keumoe, B Cichocki, M Elhabiri, B Meunier, JM Strub, M Monti, S Cianférani, SA Blandin, C Schaeffer-Reiss, E Davioud-Charvet
Proteomic Profiling of Antimalarial Plasmodione Using 3-Benz(o)ylmenadione Affinity-Based Probes Journal Article
In: Chembiochem, 2024.
Abstract | Links | BibTeX | Tags: Activity-Based Protein Profiling, Antiprotozoal agents, Biological Chemistry, blandin, Chemical Biology, M3i, Photoaffinity Labeling, proteomics
@article{I2024,
title = {Proteomic Profiling of Antimalarial Plasmodione Using 3-Benz(o)ylmenadione Affinity-Based Probes},
author = {Iacobucci I and Monaco V and Hovasse A AND Dupouy B AND Keumoe R AND Cichocki B AND Elhabiri M AND Meunier B AND Strub JM AND Monti M AND Cianférani S AND Blandin SA AND Schaeffer-Reiss C AND Davioud-Charvet E},
editor = {Wiley-VCH GmbH},
url = {https://pubmed.ncbi.nlm.nih.gov/38639212/},
doi = {10.1002/cbic.202400187},
year = {2024},
date = {2024-04-19},
journal = {Chembiochem},
abstract = {Understanding the mechanisms of drug action in malarial parasites is crucial for the development of new drugs to combat infection and to counteract drug resistance. Proteomics is a widely used approach to study host-pathogen systems and to identify drug protein targets. Plasmodione is an antiplasmodial early-lead drug exerting potent activities against young asexual and sexual blood stages in vitro with low toxicity to host cells. To elucidate its molecular mechanisms, an affinity-based protein profiling (AfBPP) approach was applied to yeast and P. falciparum proteomes. New (pro-) AfBPP probes based on the 3-benz(o)yl-6-fluoro-menadione scaffold were synthesized. With optimized conditions of both photoaffinity labeling and click reaction steps, the AfBPP protocol was then applied to a yeast proteome, yielding 11 putative drug-protein targets. Among these, we found four proteins associated with oxidoreductase activities, the hypothesized type of targets for plasmodione and its metabolites, and other proteins associated with the mitochondria. In Plasmodium parasites, the MS analysis revealed 44 potential plasmodione targets that need to be validated in further studies. Finally, the localization of a 3-benzyl-6-fluoromenadione AfBPP probe was studied in the subcellular structures of the parasite at the trophozoite stage. },
keywords = {Activity-Based Protein Profiling, Antiprotozoal agents, Biological Chemistry, blandin, Chemical Biology, M3i, Photoaffinity Labeling, proteomics},
pubstate = {published},
tppubtype = {article}
}
Understanding the mechanisms of drug action in malarial parasites is crucial for the development of new drugs to combat infection and to counteract drug resistance. Proteomics is a widely used approach to study host-pathogen systems and to identify drug protein targets. Plasmodione is an antiplasmodial early-lead drug exerting potent activities against young asexual and sexual blood stages in vitro with low toxicity to host cells. To elucidate its molecular mechanisms, an affinity-based protein profiling (AfBPP) approach was applied to yeast and P. falciparum proteomes. New (pro-) AfBPP probes based on the 3-benz(o)yl-6-fluoro-menadione scaffold were synthesized. With optimized conditions of both photoaffinity labeling and click reaction steps, the AfBPP protocol was then applied to a yeast proteome, yielding 11 putative drug-protein targets. Among these, we found four proteins associated with oxidoreductase activities, the hypothesized type of targets for plasmodione and its metabolites, and other proteins associated with the mitochondria. In Plasmodium parasites, the MS analysis revealed 44 potential plasmodione targets that need to be validated in further studies. Finally, the localization of a 3-benzyl-6-fluoromenadione AfBPP probe was studied in the subcellular structures of the parasite at the trophozoite stage.
Imler Jean-Luc, Cai Hua, Meignin Carine, Martins Nelson
Evolutionary immunology to explore original antiviral strategies Journal Article
In: Philosophical Transactions B, vol. 379, iss. 1901, 2024.
Abstract | Links | BibTeX | Tags: antiviral, imler, M3i, meignin
@article{Martins2024,
title = {Evolutionary immunology to explore original antiviral strategies},
author = {Jean-Luc Imler AND Hua Cai AND Carine Meignin AND Nelson Martins},
url = {https://royalsocietypublishing.org/doi/10.1098/rstb.2023.0068},
doi = {10.1098/rstb.2023.0068},
year = {2024},
date = {2024-03-18},
urldate = {2024-03-18},
journal = {Philosophical Transactions B},
volume = {379},
issue = {1901},
abstract = {Over the past 25 years, the field of evolutionary developmental biology (evo–devo) has used genomics and genetics to gain insight on the developmental mechanisms underlying the evolution of morphological diversity of animals. Evo–devo exploits the key insight that conserved toolkits of development (e.g. Hox genes) are used in animals to produce genetic novelties that provide adaptation to a new environment. Like development, immunity is forged by interactions with the environment, namely the microbial world. Yet, when it comes to the study of immune defence mechanisms in invertebrates, interest primarily focuses on evolutionarily conserved molecules also present in humans. Here, focusing on antiviral immunity, we argue that immune genes not conserved in humans represent an unexplored resource for the discovery of new antiviral strategies. We review recent findings on the cGAS-STING pathway and explain how cyclic dinucleotides produced by cGAS-like receptors may be used to investigate the portfolio of antiviral genes in a broad range of species. This will set the stage for evo–immuno approaches, exploiting the investment in antiviral defences made by metazoans over hundreds of millions of years of evolution.},
keywords = {antiviral, imler, M3i, meignin},
pubstate = {published},
tppubtype = {article}
}
Over the past 25 years, the field of evolutionary developmental biology (evo–devo) has used genomics and genetics to gain insight on the developmental mechanisms underlying the evolution of morphological diversity of animals. Evo–devo exploits the key insight that conserved toolkits of development (e.g. Hox genes) are used in animals to produce genetic novelties that provide adaptation to a new environment. Like development, immunity is forged by interactions with the environment, namely the microbial world. Yet, when it comes to the study of immune defence mechanisms in invertebrates, interest primarily focuses on evolutionarily conserved molecules also present in humans. Here, focusing on antiviral immunity, we argue that immune genes not conserved in humans represent an unexplored resource for the discovery of new antiviral strategies. We review recent findings on the cGAS-STING pathway and explain how cyclic dinucleotides produced by cGAS-like receptors may be used to investigate the portfolio of antiviral genes in a broad range of species. This will set the stage for evo–immuno approaches, exploiting the investment in antiviral defences made by metazoans over hundreds of millions of years of evolution.
L Hédelin, A Thiebaut, J Huang, X Li, A Lemoine, G Haas, C Meignin, H Cai, R Waterhouse, N Martins, JL Imler
Investigating the Evolution of Drosophila STING-Dependent Antiviral Innate Immunity by Multispecies Comparison of 2′3′-cGAMP Responses Journal Article
In: Molecular Biology and Evolution, vol. 41, iss. 3, 2024.
Abstract | Links | BibTeX | Tags: 2'3'-cGAMP, cGAS/STING, Drosophila, evo-immuno, imler, M3i, meignin, transcriptome
@article{JL2024,
title = {Investigating the Evolution of Drosophila STING-Dependent Antiviral Innate Immunity by Multispecies Comparison of 2′3′-cGAMP Responses},
author = {Hédelin L AND Thiebaut A AND Huang J AND Li X AND Lemoine A AND Haas G AND Meignin C AND Cai H AND Waterhouse R AND Martins N AND Imler JL},
url = {https://academic.oup.com/mbe/article/41/3/msae032/7611634},
doi = {10.1093/molbev/msae032},
year = {2024},
date = {2024-02-20},
urldate = {2024-02-20},
journal = {Molecular Biology and Evolution},
volume = {41},
issue = {3},
abstract = {Viruses represent a major threat to all animals, which defend themselves through induction of a large set of virus-stimulated genes that collectively control the infection. In vertebrates, these genes include interferons that play a critical role in the amplification of the response to infection. Virus- and interferon-stimulated genes include restriction factors targeting the different steps of the viral replication cycle, in addition to molecules associated with inflammation and adaptive immunity. Predictably, antiviral genes evolve dynamically in response to viral pressure. As a result, each animal has a unique arsenal of antiviral genes. Here, we exploit the capacity to experimentally activate the evolutionarily conserved stimulator of IFN genes (STING) signaling pathway by injection of the cyclic dinucleotide 2′3′-cyclic guanosine monophosphate-adenosine monophosphate into flies to define the repertoire of STING-regulated genes in 10 Drosophila species, spanning 40 million years of evolution. Our data reveal a set of conserved STING-regulated factors, including STING itself, a cGAS-like-receptor, the restriction factor pastel, and the antiviral protein Vago, but also 2 key components of the antiviral RNA interference pathway, Dicer-2, and Argonaute2. In addition, we identify unknown species- or lineage-specific genes that have not been previously associated with resistance to viruses. Our data provide insight into the core antiviral response in Drosophila flies and pave the way for the characterization of previously unknown antiviral effectors.},
keywords = {2'3'-cGAMP, cGAS/STING, Drosophila, evo-immuno, imler, M3i, meignin, transcriptome},
pubstate = {published},
tppubtype = {article}
}
Viruses represent a major threat to all animals, which defend themselves through induction of a large set of virus-stimulated genes that collectively control the infection. In vertebrates, these genes include interferons that play a critical role in the amplification of the response to infection. Virus- and interferon-stimulated genes include restriction factors targeting the different steps of the viral replication cycle, in addition to molecules associated with inflammation and adaptive immunity. Predictably, antiviral genes evolve dynamically in response to viral pressure. As a result, each animal has a unique arsenal of antiviral genes. Here, we exploit the capacity to experimentally activate the evolutionarily conserved stimulator of IFN genes (STING) signaling pathway by injection of the cyclic dinucleotide 2′3′-cyclic guanosine monophosphate-adenosine monophosphate into flies to define the repertoire of STING-regulated genes in 10 Drosophila species, spanning 40 million years of evolution. Our data reveal a set of conserved STING-regulated factors, including STING itself, a cGAS-like-receptor, the restriction factor pastel, and the antiviral protein Vago, but also 2 key components of the antiviral RNA interference pathway, Dicer-2, and Argonaute2. In addition, we identify unknown species- or lineage-specific genes that have not been previously associated with resistance to viruses. Our data provide insight into the core antiviral response in Drosophila flies and pave the way for the characterization of previously unknown antiviral effectors.
CF Estevez-Castro, MF Rodrigues, A Babarit, FV Ferreira, de Andrade EG, E Marois, R Cogni, ERGR Aguiar, JT Marques, RP Olmo
Neofunctionalization driven by positive selection led to the retention of the loqs2 gene encoding an Aedes specific dsRNA binding protein Journal Article
In: BMC Biol , vol. 22, no. 14, 2024.
Abstract | Links | BibTeX | Tags: Aedes mosquitoes, Double-stranded RNA (dsRNA), dsRNA binding protein (dsRBP), loqs2, M3i, marois, Marques, Olmo, RNA interference (RNAi)
@article{CF2024,
title = {Neofunctionalization driven by positive selection led to the retention of the loqs2 gene encoding an Aedes specific dsRNA binding protein},
author = {Estevez-Castro CF and Rodrigues MF and Babarit A and Ferreira FV and de Andrade EG and Marois E and Cogni R and Aguiar ERGR and Marques JT and Olmo RP},
url = {https://doi.org/10.1186/s12915-024-01821-4},
doi = {10.1186/s12915-024-01821-4},
year = {2024},
date = {2024-01-25},
urldate = {2024-01-25},
journal = {BMC Biol },
volume = {22},
number = {14},
abstract = {Background
Mosquito borne viruses, such as dengue, Zika, yellow fever and Chikungunya, cause millions of infections every year. These viruses are mostly transmitted by two urban-adapted mosquito species, Aedes aegypti and Aedes albopictus. Although mechanistic understanding remains largely unknown, Aedes mosquitoes may have unique adaptations that lower the impact of viral infection. Recently, we reported the identification of an Aedes specific double-stranded RNA binding protein (dsRBP), named Loqs2, that is involved in the control of infection by dengue and Zika viruses in mosquitoes. Preliminary analyses suggested that the loqs2 gene is a paralog of loquacious (loqs) and r2d2, two co-factors of the RNA interference (RNAi) pathway, a major antiviral mechanism in insects.
Results
Here we analyzed the origin and evolution of loqs2. Our data suggest that loqs2 originated from two independent duplications of the first double-stranded RNA binding domain of loqs that occurred before the origin of the Aedes Stegomyia subgenus, around 31 million years ago. We show that the loqs2 gene is evolving under relaxed purifying selection at a faster pace than loqs, with evidence of neofunctionalization driven by positive selection. Accordingly, we observed that Loqs2 is localized mainly in the nucleus, different from R2D2 and both isoforms of Loqs that are cytoplasmic. In contrast to r2d2 and loqs, loqs2 expression is stage- and tissue-specific, restricted mostly to reproductive tissues in adult Ae. aegypti and Ae. albopictus. Transgenic mosquitoes engineered to express loqs2 ubiquitously undergo developmental arrest at larval stages that correlates with massive dysregulation of gene expression without major effects on microRNAs or other endogenous small RNAs, classically associated with RNA interference.
Conclusions
Our results uncover the peculiar origin and neofunctionalization of loqs2 driven by positive selection. This study shows an example of unique adaptations in Aedes mosquitoes that could ultimately help explain their effectiveness as virus vectors.},
keywords = {Aedes mosquitoes, Double-stranded RNA (dsRNA), dsRNA binding protein (dsRBP), loqs2, M3i, marois, Marques, Olmo, RNA interference (RNAi)},
pubstate = {published},
tppubtype = {article}
}
Background
Mosquito borne viruses, such as dengue, Zika, yellow fever and Chikungunya, cause millions of infections every year. These viruses are mostly transmitted by two urban-adapted mosquito species, Aedes aegypti and Aedes albopictus. Although mechanistic understanding remains largely unknown, Aedes mosquitoes may have unique adaptations that lower the impact of viral infection. Recently, we reported the identification of an Aedes specific double-stranded RNA binding protein (dsRBP), named Loqs2, that is involved in the control of infection by dengue and Zika viruses in mosquitoes. Preliminary analyses suggested that the loqs2 gene is a paralog of loquacious (loqs) and r2d2, two co-factors of the RNA interference (RNAi) pathway, a major antiviral mechanism in insects.
Results
Here we analyzed the origin and evolution of loqs2. Our data suggest that loqs2 originated from two independent duplications of the first double-stranded RNA binding domain of loqs that occurred before the origin of the Aedes Stegomyia subgenus, around 31 million years ago. We show that the loqs2 gene is evolving under relaxed purifying selection at a faster pace than loqs, with evidence of neofunctionalization driven by positive selection. Accordingly, we observed that Loqs2 is localized mainly in the nucleus, different from R2D2 and both isoforms of Loqs that are cytoplasmic. In contrast to r2d2 and loqs, loqs2 expression is stage- and tissue-specific, restricted mostly to reproductive tissues in adult Ae. aegypti and Ae. albopictus. Transgenic mosquitoes engineered to express loqs2 ubiquitously undergo developmental arrest at larval stages that correlates with massive dysregulation of gene expression without major effects on microRNAs or other endogenous small RNAs, classically associated with RNA interference.
Conclusions
Our results uncover the peculiar origin and neofunctionalization of loqs2 driven by positive selection. This study shows an example of unique adaptations in Aedes mosquitoes that could ultimately help explain their effectiveness as virus vectors.
Mosquito borne viruses, such as dengue, Zika, yellow fever and Chikungunya, cause millions of infections every year. These viruses are mostly transmitted by two urban-adapted mosquito species, Aedes aegypti and Aedes albopictus. Although mechanistic understanding remains largely unknown, Aedes mosquitoes may have unique adaptations that lower the impact of viral infection. Recently, we reported the identification of an Aedes specific double-stranded RNA binding protein (dsRBP), named Loqs2, that is involved in the control of infection by dengue and Zika viruses in mosquitoes. Preliminary analyses suggested that the loqs2 gene is a paralog of loquacious (loqs) and r2d2, two co-factors of the RNA interference (RNAi) pathway, a major antiviral mechanism in insects.
Results
Here we analyzed the origin and evolution of loqs2. Our data suggest that loqs2 originated from two independent duplications of the first double-stranded RNA binding domain of loqs that occurred before the origin of the Aedes Stegomyia subgenus, around 31 million years ago. We show that the loqs2 gene is evolving under relaxed purifying selection at a faster pace than loqs, with evidence of neofunctionalization driven by positive selection. Accordingly, we observed that Loqs2 is localized mainly in the nucleus, different from R2D2 and both isoforms of Loqs that are cytoplasmic. In contrast to r2d2 and loqs, loqs2 expression is stage- and tissue-specific, restricted mostly to reproductive tissues in adult Ae. aegypti and Ae. albopictus. Transgenic mosquitoes engineered to express loqs2 ubiquitously undergo developmental arrest at larval stages that correlates with massive dysregulation of gene expression without major effects on microRNAs or other endogenous small RNAs, classically associated with RNA interference.
Conclusions
Our results uncover the peculiar origin and neofunctionalization of loqs2 driven by positive selection. This study shows an example of unique adaptations in Aedes mosquitoes that could ultimately help explain their effectiveness as virus vectors.
Green Emily I, Jaouen Etienne, Klug Dennis, Olmo Roenick Proveti, Gautier Amandine, Blandin Stéphanie, Marois Eric
A population modification gene drive targeting both Saglin and Lipophorin disables Plasmodium transmission in Anopheles mosquitoes Journal Article
In: Genetics and Genomics, 2024.
Abstract | Links | BibTeX | Tags: Anopheles, blandin, gene drive, Lipophorin, M3i, marois, marque, Marques, Olmo, Plasmodium, Saglin
@article{Green2024,
title = {A population modification gene drive targeting both Saglin and Lipophorin disables Plasmodium transmission in Anopheles mosquitoes},
author = {Emily I Green and Etienne Jaouen and Dennis Klug and Roenick Proveti Olmo and Amandine Gautier and Stéphanie Blandin and Eric Marois},
url = {https://elifesciences.org/articles/93142},
doi = {10.7554/eLife.93142},
year = {2024},
date = {2024-01-12},
journal = {Genetics and Genomics},
abstract = {Lipophorin is an essential, highly expressed lipid transport protein that is secreted and circulates in insect hemolymph. We hijacked the Anopheles coluzzii Lipophorin gene to make it co-express a single-chain version of antibody 2A10, which binds sporozoites of the malaria parasite Plasmodium falciparum. The resulting transgenic mosquitoes show a markedly decreased ability to transmit Plasmodium berghei expressing the P. falciparum circumsporozoite protein to mice. To force the spread of this antimalarial transgene in a mosquito population, we designed and tested several CRISPR/Cas9-based gene drives. One of these is installed in, and disrupts, the pro-parasitic gene Saglin and also cleaves wild-type Lipophorin, causing the anti-malarial modified Lipophorin version to replace the wild type and hitch-hike together with the Saglin drive. Although generating drive-resistant alleles and showing instability in its gRNA-encoding multiplex array, the Saglin-based gene drive reached high levels in caged mosquito populations and efficiently promoted the simultaneous spread of the antimalarial Lipophorin::Sc2A10 allele. This combination is expected to decrease parasite transmission via two different mechanisms. This work contributes to the design of novel strategies to spread antimalarial transgenes in mosquitoes, and illustrates some expected and unexpected outcomes encountered when establishing a population modification gene drive.},
keywords = {Anopheles, blandin, gene drive, Lipophorin, M3i, marois, marque, Marques, Olmo, Plasmodium, Saglin},
pubstate = {published},
tppubtype = {article}
}
Lipophorin is an essential, highly expressed lipid transport protein that is secreted and circulates in insect hemolymph. We hijacked the Anopheles coluzzii Lipophorin gene to make it co-express a single-chain version of antibody 2A10, which binds sporozoites of the malaria parasite Plasmodium falciparum. The resulting transgenic mosquitoes show a markedly decreased ability to transmit Plasmodium berghei expressing the P. falciparum circumsporozoite protein to mice. To force the spread of this antimalarial transgene in a mosquito population, we designed and tested several CRISPR/Cas9-based gene drives. One of these is installed in, and disrupts, the pro-parasitic gene Saglin and also cleaves wild-type Lipophorin, causing the anti-malarial modified Lipophorin version to replace the wild type and hitch-hike together with the Saglin drive. Although generating drive-resistant alleles and showing instability in its gRNA-encoding multiplex array, the Saglin-based gene drive reached high levels in caged mosquito populations and efficiently promoted the simultaneous spread of the antimalarial Lipophorin::Sc2A10 allele. This combination is expected to decrease parasite transmission via two different mechanisms. This work contributes to the design of novel strategies to spread antimalarial transgenes in mosquitoes, and illustrates some expected and unexpected outcomes encountered when establishing a population modification gene drive.
Marois Eric
Using the CRISPR / Cas9 system for genome editing in Anopheles mosquitoes Online
2024, visited: 08.01.2024.
Abstract | Links | BibTeX | Tags: Anopheles, CRISPR/Cas9, Genome editing, M3i, marois, mosquitoes
@online{Marois2024,
title = {Using the CRISPR / Cas9 system for genome editing in Anopheles mosquitoes},
author = { Eric Marois},
url = {https://hal.science/hal-04380430/document},
doi = {HAL Id: hal-04380430},
year = {2024},
date = {2024-01-08},
urldate = {2024-01-08},
journal = {HAL science},
abstract = {The advent of the CRISPR / Cas9 technology permits the targeted editing of mosquito genomes, ranging from site-directed mutagenesis of genes of interest yielding knockout mutations (which arise by insertion / deletion of a few nucleotides) to site-specific insertion of exogenous DNA sequences such as fluorescence markers or even large gene drive cassettes, themselves encoding the components of the CRISPR / Cas9 system. To obtain these heritable targeted changes, genome editing requires the delivery of Cas9 protein and its guide RNA(s) to the developing germ tissue of an embryo. Different species require adaptation of this basic principle to accommodate for their specific biology. Here, we describe a technical pipeline based on delivering the CRISPR/Cas9 components in the form of injected plasmid or as transgenes, resulting in highly efficient gene editing in Anopheles malaria vector mosquitoes. We have reliably employed these methods to mutagenize > 20 different loci of interest in Anopheles coluzzii to date. },
keywords = {Anopheles, CRISPR/Cas9, Genome editing, M3i, marois, mosquitoes},
pubstate = {published},
tppubtype = {online}
}
The advent of the CRISPR / Cas9 technology permits the targeted editing of mosquito genomes, ranging from site-directed mutagenesis of genes of interest yielding knockout mutations (which arise by insertion / deletion of a few nucleotides) to site-specific insertion of exogenous DNA sequences such as fluorescence markers or even large gene drive cassettes, themselves encoding the components of the CRISPR / Cas9 system. To obtain these heritable targeted changes, genome editing requires the delivery of Cas9 protein and its guide RNA(s) to the developing germ tissue of an embryo. Different species require adaptation of this basic principle to accommodate for their specific biology. Here, we describe a technical pipeline based on delivering the CRISPR/Cas9 components in the form of injected plasmid or as transgenes, resulting in highly efficient gene editing in Anopheles malaria vector mosquitoes. We have reliably employed these methods to mutagenize > 20 different loci of interest in Anopheles coluzzii to date.
2023
A Hinze, J Pelletier, M Ghaninia, E Marois, Hill, R S Ignell
Knockout of OR39 reveals redundancy in the olfactory pathway regulating the acquisition of host seeking in Anopheles coluzzii Journal Article
In: Proc Biol Sci 290(2011):20232092, vol. 290, no. 2011, 2023.
Abstract | Links | BibTeX | Tags: CRISPR–Cas9, human odour, M3i, marois, mosquito flight, odour valence, SSR
@article{Hinze2023,
title = {Knockout of OR39 reveals redundancy in the olfactory pathway regulating the acquisition of host seeking in Anopheles coluzzii},
author = {Hinze A and Pelletier J and Ghaninia M and Marois E and Hill and S Ignell R},
url = {https://doi.org/10.1098/rspb.2023.2092},
doi = {10.1098/rspb.2023.2092},
year = {2023},
date = {2023-11-29},
urldate = {2023-11-29},
booktitle = {Proc Biol Sci},
journal = {Proc Biol Sci 290(2011):20232092},
volume = {290},
number = {2011},
abstract = {The attraction of anthropophilic mosquitoes to human host cues, such as body odour and carbon dioxide, gradually increases during adult maturation. This acquisition of host-seeking behaviour correlates with age-dependent changes in odorant receptor (OR) transcript abundance and sensitivity of olfactory sensory neurons (OSNs). One OR gene of the human malaria vector, Anopheles coluzzii, AcolOR39, is significantly downregulated in mature females, and a cognate ligand of AcolOR39, sulcatone, a major component of human emanations, mediates the observed behavioural inhibition of newly emerged (teneral) females to human body odour. Knockout of AcolOR39, using CRISPR–Cas9 mutagenesis, selectively abolished sulcatone detection in OSNs, housed in trichoid sensilla. However, knockout of AcolOR39 altered neither the response rate nor the flight behaviour of teneral females in a wind tunnel, indicating the involvement of other genes, and thus a redundancy, in regulating the acquisition of host seeking in mosquitoes.},
keywords = {CRISPR–Cas9, human odour, M3i, marois, mosquito flight, odour valence, SSR},
pubstate = {published},
tppubtype = {article}
}
The attraction of anthropophilic mosquitoes to human host cues, such as body odour and carbon dioxide, gradually increases during adult maturation. This acquisition of host-seeking behaviour correlates with age-dependent changes in odorant receptor (OR) transcript abundance and sensitivity of olfactory sensory neurons (OSNs). One OR gene of the human malaria vector, Anopheles coluzzii, AcolOR39, is significantly downregulated in mature females, and a cognate ligand of AcolOR39, sulcatone, a major component of human emanations, mediates the observed behavioural inhibition of newly emerged (teneral) females to human body odour. Knockout of AcolOR39, using CRISPR–Cas9 mutagenesis, selectively abolished sulcatone detection in OSNs, housed in trichoid sensilla. However, knockout of AcolOR39 altered neither the response rate nor the flight behaviour of teneral females in a wind tunnel, indicating the involvement of other genes, and thus a redundancy, in regulating the acquisition of host seeking in mosquitoes.
SC Weng, I Antoshechkin, E Marois, O Akbari
Efficient Sex Separation by Exploiting Differential Alternative Splicing of a Dominant Marker in Aedes aegypti Journal Article
In: PLoS Genetics, 2023.
Abstract | Links | BibTeX | Tags: Aedes aegypti, Introns, larvae, M3i, marois, mosquitoes, pupae, rna sequencing, transcriptome analyses
@article{nokey,
title = {Efficient Sex Separation by Exploiting Differential Alternative Splicing of a Dominant Marker in Aedes aegypti},
author = {Weng SC and Antoshechkin I and Marois E and Akbari O},
url = {https://doi.org/10.1371/journal.pgen.1011065},
doi = {10.1371/journal.pgen.1011065},
year = {2023},
date = {2023-11-27},
urldate = {2023-11-27},
journal = {PLoS Genetics},
abstract = {Only female mosquitoes consume blood giving them the opportunity to transmit deadly human pathogens. Therefore, it is critical to remove females before conducting releases for genetic biocontrol interventions. Here we describe a robust sex-sorting approach termed SEPARATOR (Sexing Element Produced by Alternative RNA-splicing of A Transgenic Observable Reporter) that exploits sex-specific alternative splicing of an innocuous reporter to ensure exclusive dominant male-specific expression. Using SEPARATOR, we demonstrate reliable sex selection from early larval and pupal stages in Aedes aegypti, and use a Complex Object Parametric Analyzer and Sorter (COPAS) to demonstrate scalable high-throughput sex-selection of first instar larvae. Additionally, we use this approach to sequence the transcriptomes of early larval males and females and find several genes that are sex-specifically expressed. SEPARATOR can simplify mass production of males for release programs and is designed to be cross-species portable and should be instrumental for genetic biocontrol interventions.},
keywords = {Aedes aegypti, Introns, larvae, M3i, marois, mosquitoes, pupae, rna sequencing, transcriptome analyses},
pubstate = {published},
tppubtype = {article}
}
Only female mosquitoes consume blood giving them the opportunity to transmit deadly human pathogens. Therefore, it is critical to remove females before conducting releases for genetic biocontrol interventions. Here we describe a robust sex-sorting approach termed SEPARATOR (Sexing Element Produced by Alternative RNA-splicing of A Transgenic Observable Reporter) that exploits sex-specific alternative splicing of an innocuous reporter to ensure exclusive dominant male-specific expression. Using SEPARATOR, we demonstrate reliable sex selection from early larval and pupal stages in Aedes aegypti, and use a Complex Object Parametric Analyzer and Sorter (COPAS) to demonstrate scalable high-throughput sex-selection of first instar larvae. Additionally, we use this approach to sequence the transcriptomes of early larval males and females and find several genes that are sex-specifically expressed. SEPARATOR can simplify mass production of males for release programs and is designed to be cross-species portable and should be instrumental for genetic biocontrol interventions.
AI Kalita, E Marois, M Kozielska, FJ Weissing, E Jaouen, Möckel MM and Rühle F, F Butter, MF Basilicata, CI Keller Valsecchi
The sex-specific factor SOA controls dosage compensation in Anopheles mosquitoes Journal Article
In: Nature, vol. 623, pp. 175–182, 2023.
Abstract | Links | BibTeX | Tags: Anopheles, M3i, marois, SOA
@article{AI2023,
title = {The sex-specific factor SOA controls dosage compensation in Anopheles mosquitoes},
author = {Kalita AI and Marois E and Kozielska M and Weissing FJ and Jaouen E and Möckel MM and, Rühle F and Butter F and Basilicata MF and Keller Valsecchi CI},
url = {https://www.nature.com/articles/s41586-023-06641-0},
doi = {10.1038/s41586-023-06641-0.},
year = {2023},
date = {2023-09-28},
journal = {Nature},
volume = {623},
pages = {175–182},
abstract = {The Anopheles mosquito is one of thousands of species in which sex differences play a central part in their biology, as only females need a blood meal to produce eggs. Sex differentiation is regulated by sex chromosomes, but their presence creates a dosage imbalance between males (XY) and females (XX). Dosage compensation (DC) can re-equilibrate the expression of sex chromosomal genes. However, because DC mechanisms have only been fully characterized in a few model organisms, key questions about its evolutionary diversity and functional necessity remain unresolved1. Here we report the discovery of a previously uncharacterized gene (sex chromosome activation (SOA)) as a master regulator of DC in the malaria mosquito Anopheles gambiae. Sex-specific alternative splicing prevents functional SOA protein expression in females. The male isoform encodes a DNA-binding protein that binds the promoters of active X chromosomal genes. Expressing male SOA is sufficient to induce DC in female cells. Male mosquitoes lacking SOA or female mosquitoes ectopically expressing the male isoform exhibit X chromosome misregulation, which is compatible with viability but causes developmental delay. Thus, our molecular analyses of a DC master regulator in a non-model organism elucidates the evolutionary steps that lead to the establishment of a chromosome-specific fine-tuning mechanism.},
keywords = {Anopheles, M3i, marois, SOA},
pubstate = {published},
tppubtype = {article}
}
The Anopheles mosquito is one of thousands of species in which sex differences play a central part in their biology, as only females need a blood meal to produce eggs. Sex differentiation is regulated by sex chromosomes, but their presence creates a dosage imbalance between males (XY) and females (XX). Dosage compensation (DC) can re-equilibrate the expression of sex chromosomal genes. However, because DC mechanisms have only been fully characterized in a few model organisms, key questions about its evolutionary diversity and functional necessity remain unresolved1. Here we report the discovery of a previously uncharacterized gene (sex chromosome activation (SOA)) as a master regulator of DC in the malaria mosquito Anopheles gambiae. Sex-specific alternative splicing prevents functional SOA protein expression in females. The male isoform encodes a DNA-binding protein that binds the promoters of active X chromosomal genes. Expressing male SOA is sufficient to induce DC in female cells. Male mosquitoes lacking SOA or female mosquitoes ectopically expressing the male isoform exhibit X chromosome misregulation, which is compatible with viability but causes developmental delay. Thus, our molecular analyses of a DC master regulator in a non-model organism elucidates the evolutionary steps that lead to the establishment of a chromosome-specific fine-tuning mechanism.
SR Abbo, de Almeida JPP, RP Olmo, C Balvers, JS Griep, C Linthout, CJM Koenraadt, BM Silva, JJ Fros, ERGR Aguiar, E Marois, GP Pijlman, JT Marques
The virome of the invasive Asian bush mosquito Aedes japonicus in Europe Journal Article
In: Virus Evol., vol. 9, iss. 2, 2023.
Abstract | Links | BibTeX | Tags: Aedes japonicus, anphevirus, bunyavirus, M3i, marois, Marques, metagenomics, mosquito, Olmo, rhabdovirus, RNA Interference, totivirus, virome
@article{Abbo2023,
title = {The virome of the invasive Asian bush mosquito Aedes japonicus in Europe},
author = {Abbo SR and de Almeida JPP and Olmo RP and Balvers C and Griep JS and Linthout C and Koenraadt CJM and Silva BM and Fros JJ and Aguiar ERGR and Marois E and Pijlman GP and Marques JT},
doi = {10.1093/ve/vead041},
year = {2023},
date = {2023-09-22},
urldate = {2023-09-22},
journal = {Virus Evol.},
volume = {9},
issue = {2},
abstract = {The Asian bush mosquito Aedes japonicus is rapidly invading North America and Europe. Due to its potential to transmit multiple pathogenic arthropod-borne (arbo)viruses including Zika virus, West Nile virus, and chikungunya virus, it is important to understand the biology of this vector mosquito in more detail. In addition to arboviruses, mosquitoes can also carry insect-specific viruses that are receiving increasing attention due to their potential effects on host physiology and arbovirus transmission. In this study, we characterized the collection of viruses, referred to as the virome, circulating in Ae. japonicus populations in the Netherlands and France. Applying a small RNA-based metagenomic approach to Ae. japonicus, we uncovered a distinct group of viruses present in samples from both the Netherlands and France. These included one known virus, Ae. japonicus narnavirus 1 (AejapNV1), and three new virus species that we named Ae. japonicus totivirus 1 (AejapTV1), Ae. japonicus anphevirus 1 (AejapAV1) and Ae. japonicus bunyavirus 1 (AejapBV1). We also discovered sequences that were presumably derived from two additional novel viruses: Ae. japonicus bunyavirus 2 (AejapBV2) and Ae. japonicus rhabdovirus 1 (AejapRV1). All six viruses induced strong RNA interference responses, including the production of twenty-one nucleotide-sized small interfering RNAs, a signature of active replication in the host. Notably, AejapBV1 and AejapBV2 belong to different viral families; however, no RNA-dependent RNA polymerase sequence has been found for AejapBV2. Intriguingly, our small RNA-based approach identified an ∼1-kb long ambigrammatic RNA that is associated with AejapNV1 as a secondary segment but showed no similarity to any sequence in public databases. We confirmed the presence of AejapNV1 primary and secondary segments, AejapTV1, AejapAV1, and AejapBV1 by reverse transcriptase polymerase chain reaction (PCR) in wild-caught Ae. japonicus mosquitoes. AejapNV1 and AejapTV1 were found at high prevalence (87-100 per cent) in adult females, adult males, and larvae. Using a small RNA-based, sequence-independent metagenomic strategy, we uncovered a conserved and prevalent virome among Ae. japonicus mosquito populations. The high prevalence of AejapNV1 and AejapTV1 across all tested mosquito life stages suggests that these viruses are intimately associated with Ae. japonicus.},
keywords = {Aedes japonicus, anphevirus, bunyavirus, M3i, marois, Marques, metagenomics, mosquito, Olmo, rhabdovirus, RNA Interference, totivirus, virome},
pubstate = {published},
tppubtype = {article}
}
The Asian bush mosquito Aedes japonicus is rapidly invading North America and Europe. Due to its potential to transmit multiple pathogenic arthropod-borne (arbo)viruses including Zika virus, West Nile virus, and chikungunya virus, it is important to understand the biology of this vector mosquito in more detail. In addition to arboviruses, mosquitoes can also carry insect-specific viruses that are receiving increasing attention due to their potential effects on host physiology and arbovirus transmission. In this study, we characterized the collection of viruses, referred to as the virome, circulating in Ae. japonicus populations in the Netherlands and France. Applying a small RNA-based metagenomic approach to Ae. japonicus, we uncovered a distinct group of viruses present in samples from both the Netherlands and France. These included one known virus, Ae. japonicus narnavirus 1 (AejapNV1), and three new virus species that we named Ae. japonicus totivirus 1 (AejapTV1), Ae. japonicus anphevirus 1 (AejapAV1) and Ae. japonicus bunyavirus 1 (AejapBV1). We also discovered sequences that were presumably derived from two additional novel viruses: Ae. japonicus bunyavirus 2 (AejapBV2) and Ae. japonicus rhabdovirus 1 (AejapRV1). All six viruses induced strong RNA interference responses, including the production of twenty-one nucleotide-sized small interfering RNAs, a signature of active replication in the host. Notably, AejapBV1 and AejapBV2 belong to different viral families; however, no RNA-dependent RNA polymerase sequence has been found for AejapBV2. Intriguingly, our small RNA-based approach identified an ∼1-kb long ambigrammatic RNA that is associated with AejapNV1 as a secondary segment but showed no similarity to any sequence in public databases. We confirmed the presence of AejapNV1 primary and secondary segments, AejapTV1, AejapAV1, and AejapBV1 by reverse transcriptase polymerase chain reaction (PCR) in wild-caught Ae. japonicus mosquitoes. AejapNV1 and AejapTV1 were found at high prevalence (87-100 per cent) in adult females, adult males, and larvae. Using a small RNA-based, sequence-independent metagenomic strategy, we uncovered a conserved and prevalent virome among Ae. japonicus mosquito populations. The high prevalence of AejapNV1 and AejapTV1 across all tested mosquito life stages suggests that these viruses are intimately associated with Ae. japonicus.
H Cai, L Li, KM Slavik, J Huang, T Yin, X Ai, L Hédelin, G Haas, Z Xiang, Y Yang, X Li, Y Chen, Z Wei, H Deng, D Chen, R Jiao, N Martins, C Meignin, PJ Kranzusch, JL Imler
The virus-induced cyclic dinucleotide 2'3'-c-di-GMP mediates STING-dependent antiviral immunity in Drosophila Journal Article
In: Immunity, vol. 56, iss. 9, pp. 1991-2005, 2023.
Abstract | Links | BibTeX | Tags: c-di-GMP, cGAMP, cGAS, cGLR, cyclic dinucleotide, Drosophila, Evolution, Hua, imler, M3i, meignin, pattern recognition receptor, STING, virus
@article{nokey,
title = {The virus-induced cyclic dinucleotide 2'3'-c-di-GMP mediates STING-dependent antiviral immunity in Drosophila},
author = {Cai H and Li L and Slavik KM and Huang J and Yin T and Ai X and Hédelin L and Haas G and Xiang Z and Yang Y and Li X and Chen Y and Wei Z and Deng H and Chen D and Jiao R and Martins N and Meignin C and Kranzusch PJ and Imler JL},
editor = {Elsevier Inc. },
url = {https://pubmed.ncbi.nlm.nih.gov/37659413/},
doi = {10.1016/j.immuni.2023.08.006 },
year = {2023},
date = {2023-09-12},
urldate = {2023-09-12},
journal = {Immunity},
volume = {56},
issue = {9},
pages = {1991-2005},
abstract = {In mammals, the enzyme cGAS senses the presence of cytosolic DNA and synthesizes the cyclic dinucleotide (CDN) 2'3'-cGAMP, which triggers STING-dependent immunity. In Drosophila melanogaster, two cGAS-like receptors (cGLRs) produce 3'2'-cGAMP and 2'3'-cGAMP to activate STING. We explored CDN-mediated immunity in 14 Drosophila species covering 50 million years of evolution and found that 2'3'-cGAMP and 3'2'-cGAMP failed to control infection by Drosophila C virus in D. serrata and two other species. We discovered diverse CDNs produced in a cGLR-dependent manner in response to viral infection in D. melanogaster, including 2'3'-c-di-GMP. This CDN was a more potent STING agonist than cGAMP in D. melanogaster and it also activated a strong antiviral transcriptional response in D. serrata. Our results shed light on the evolution of cGLRs in flies and provide a basis for understanding the function and regulation of this emerging family of pattern recognition receptors in animal innate immunity. },
keywords = {c-di-GMP, cGAMP, cGAS, cGLR, cyclic dinucleotide, Drosophila, Evolution, Hua, imler, M3i, meignin, pattern recognition receptor, STING, virus},
pubstate = {published},
tppubtype = {article}
}
In mammals, the enzyme cGAS senses the presence of cytosolic DNA and synthesizes the cyclic dinucleotide (CDN) 2'3'-cGAMP, which triggers STING-dependent immunity. In Drosophila melanogaster, two cGAS-like receptors (cGLRs) produce 3'2'-cGAMP and 2'3'-cGAMP to activate STING. We explored CDN-mediated immunity in 14 Drosophila species covering 50 million years of evolution and found that 2'3'-cGAMP and 3'2'-cGAMP failed to control infection by Drosophila C virus in D. serrata and two other species. We discovered diverse CDNs produced in a cGLR-dependent manner in response to viral infection in D. melanogaster, including 2'3'-c-di-GMP. This CDN was a more potent STING agonist than cGAMP in D. melanogaster and it also activated a strong antiviral transcriptional response in D. serrata. Our results shed light on the evolution of cGLRs in flies and provide a basis for understanding the function and regulation of this emerging family of pattern recognition receptors in animal innate immunity.
Marois Eric
Screening Mosquito Larvae Under a Fluorescence Binocular Microscope Journal Article
In: Cold Spring Harb Protoc, 2023.
Links | BibTeX | Tags: fluorescent protein, M3i, marois
@article{Marois2023,
title = {Screening Mosquito Larvae Under a Fluorescence Binocular Microscope},
author = {Eric Marois},
doi = {10.1101/pdb.prot108306 },
year = {2023},
date = {2023-09-11},
urldate = {2023-09-11},
journal = {Cold Spring Harb Protoc},
keywords = {fluorescent protein, M3i, marois},
pubstate = {published},
tppubtype = {article}
}
PELLETIER Julien, DAWIT Mengistu, GHANINIA Majid, MAROIS Eric, IGNELL Rickard
A mosquito-specific antennal protein is critical for the attraction to human odor in the malaria vector Anopheles gambiae Journal Article
In: Insect Biochemistry and Molecular Biology, vol. 159, iss. August 2023, 2023.
Abstract | Links | BibTeX | Tags: antenna, chemoreceptor, M3i, marois, mosquitoes, olfaction
@article{IGNELL2023,
title = {A mosquito-specific antennal protein is critical for the attraction to human odor in the malaria vector Anopheles gambiae},
author = {Julien PELLETIER AND Mengistu DAWIT AND Majid GHANINIA AND Eric MAROIS AND Rickard IGNELL},
editor = { },
url = {https://doi.org/10.1016/j.ibmb.2023.103988},
doi = {j.ibmb.2023.103988},
year = {2023},
date = {2023-07-11},
urldate = {2023-07-11},
journal = {Insect Biochemistry and Molecular Biology},
volume = {159},
issue = {August 2023},
abstract = {Mosquitoes rely mainly on the sense of smell to decipher their environment and locate suitable food sources, hosts for blood feeding and oviposition sites. The molecular bases of olfaction involve multigenic families of olfactory proteins that have evolved to interact with a narrow set of odorants that are critical for survival. Understanding the complex interplay between diversified repertoires of olfactory proteins and ecologically-relevant odorant signals, which elicit important behaviors, is fundamental for the design of novel control strategies targeting the sense of smell of disease vector mosquitoes. Previously, large multigene families of odorant receptor and ionotropic receptor proteins, as well as a subset of odorant-binding proteins have been shown to mediate the selectivity and sensitivity of the mosquito olfactory system. In this study, we identify a mosquito-specific antennal protein (MSAP) gene as a novel molecular actor of odorant reception. MSAP is highly conserved across mosquito species and is transcribed at an extremely high level in female antennae. In order to understand its role in the mosquito olfactory system, we generated knockout mutant lines in Anopheles gambiae, and performed comparative analysis of behavioral and physiological responses to human-associated odorants. We found that MSAP promotes female mosquito attraction to human odor and enhances the sensitivity of the antennae to a variety of odorants. These findings suggest that MSAP is an important component of the mosquito olfactory system, which until now has gone completely unnoticed.},
keywords = {antenna, chemoreceptor, M3i, marois, mosquitoes, olfaction},
pubstate = {published},
tppubtype = {article}
}
Mosquitoes rely mainly on the sense of smell to decipher their environment and locate suitable food sources, hosts for blood feeding and oviposition sites. The molecular bases of olfaction involve multigenic families of olfactory proteins that have evolved to interact with a narrow set of odorants that are critical for survival. Understanding the complex interplay between diversified repertoires of olfactory proteins and ecologically-relevant odorant signals, which elicit important behaviors, is fundamental for the design of novel control strategies targeting the sense of smell of disease vector mosquitoes. Previously, large multigene families of odorant receptor and ionotropic receptor proteins, as well as a subset of odorant-binding proteins have been shown to mediate the selectivity and sensitivity of the mosquito olfactory system. In this study, we identify a mosquito-specific antennal protein (MSAP) gene as a novel molecular actor of odorant reception. MSAP is highly conserved across mosquito species and is transcribed at an extremely high level in female antennae. In order to understand its role in the mosquito olfactory system, we generated knockout mutant lines in Anopheles gambiae, and performed comparative analysis of behavioral and physiological responses to human-associated odorants. We found that MSAP promotes female mosquito attraction to human odor and enhances the sensitivity of the antennae to a variety of odorants. These findings suggest that MSAP is an important component of the mosquito olfactory system, which until now has gone completely unnoticed.
C Lutrat, M Burckbuchler, RP Olmo, R Beugnon, A Fontaine, OS Akbari, R Argiles-Herrero, T Baldet, J Bouyer, E Marois
Combining two Genetic Sexing Strains allows sorting of non-transgenic males for Aedes genetic control Journal Article
In: Commun Biol., vol. 6, iss. 1, pp. 646, 2023.
Abstract | Links | BibTeX | Tags: Aedes, M3i, marois, mosquitoes, Olmo, vectoring
@article{Lutrat2023,
title = {Combining two Genetic Sexing Strains allows sorting of non-transgenic males for Aedes genetic control},
author = {Lutrat C and Burckbuchler M and Olmo RP and Beugnon R and Fontaine A and Akbari OS and Argiles-Herrero R and Baldet T and Bouyer J and Marois E},
url = {https://www.nature.com/articles/s42003-023-05030-7},
doi = {10.1038/s42003-023-05030-7},
year = {2023},
date = {2023-06-16},
urldate = {2023-06-16},
journal = {Commun Biol.},
volume = {6},
issue = {1},
pages = {646},
abstract = {Chemical control of disease vectoring mosquitoes Aedes albopictus and Aedes aegypti is costly, unsustainable, and increasingly ineffective due to the spread of insecticide resistance. The Sterile Insect Technique is a valuable alternative but is limited by slow, error-prone, and wasteful sex-separation methods. Here, we present four Genetic Sexing Strains (two for each Aedes species) based on fluorescence markers linked to the m and M sex loci, allowing for the isolation of transgenic males. Furthermore, we demonstrate how combining these sexing strains enables the production of non-transgenic males. In a mass-rearing facility, 100,000 first instar male larvae could be sorted in under 1.5 h with an estimated 0.01–0.1% female contamination on a single machine. Cost-efficiency analyses revealed that using these strains could result in important savings while setting up and running a mass-rearing facility. Altogether, these Genetic Sexing Strains should enable a major upscaling in control programmes against these important vectors.},
keywords = {Aedes, M3i, marois, mosquitoes, Olmo, vectoring},
pubstate = {published},
tppubtype = {article}
}
Chemical control of disease vectoring mosquitoes Aedes albopictus and Aedes aegypti is costly, unsustainable, and increasingly ineffective due to the spread of insecticide resistance. The Sterile Insect Technique is a valuable alternative but is limited by slow, error-prone, and wasteful sex-separation methods. Here, we present four Genetic Sexing Strains (two for each Aedes species) based on fluorescence markers linked to the m and M sex loci, allowing for the isolation of transgenic males. Furthermore, we demonstrate how combining these sexing strains enables the production of non-transgenic males. In a mass-rearing facility, 100,000 first instar male larvae could be sorted in under 1.5 h with an estimated 0.01–0.1% female contamination on a single machine. Cost-efficiency analyses revealed that using these strains could result in important savings while setting up and running a mass-rearing facility. Altogether, these Genetic Sexing Strains should enable a major upscaling in control programmes against these important vectors.
Labbé Frédéric, Abdeladhim Maha, Abrudan Jenica, Araki Alejandra Saori, Araujo Ricardo N., Arensburger Peter, Benoit Joshua B., Brazil Reginaldo Pecanha, Bruno Rafaela V., da Silva Rivas Gustavo Bueno, de Abreu Vinicius Carvalho, Charamis Jason, Coutinho-Abreu Iliano V., da Costa-Latgé Samara G., Darby Alistair, Dillon Viv M., Emrich Scott J., Fernandez-Medina Daniela, Gontijo Nelder Figueiredo, Flanley Catherine M., Gatherer Derek, Genta Fernando A., Gesing Sandra, Giraldo-Calderón Gloria I., Gomes Bruno, Aguiar Eric Roberto Guimaraes Rocha, Hamilton James G. C., Hamarsheh Omar, Hawksworth Mallory, Hendershot Jacob M., Hickner Paul V., Imler Jean-Luc, Ioannidis Panagiotis, Jennings Emily C., Kamhawi Shaden, Karageorgiou Charikleia, Kennedy Ryan C., Krueger Andreas, Latorre-Estivalis José M., Ligoxygakis Petros, Meireles-Filho Antonio Carlos A., Minx Patrick, Miranda Jose Carlos, Montague Michael J., Nowling Ronald J., Oliveira Fabiano, Ortigão-Farias João, Pavan Marcio G., Pereira Marcos Horacio, Pitaluga Andre Nobrega, Olmo Roenick Proveti, Ramalho-Ortigao Marcelo, Ribeiro José M. C., Rosendale Andrew J., Sant’Anna Mauricio R. V., Scherer Steven E., Secundino Nágila F. C., Shoue Douglas A., da Silva Moraes Caroline, Gesto João Silveira Moledo, Souza Nataly Araujo, Syed Zainulabueddin, Tadros Samuel, Teles-de-Freitas Rayane, Telleria Erich L., Tomlinson Chad, Traub-Csekö Yara M., Marques João Trindade, Tu Zhijian, Unger Maria F., Valenzuela Jesus, Ferreira Flávia V., de Oliveira Karla P. V., Vigoder Felipe M., Vontas John, Wang Lihui, Weedall Gareth D., Zhioua Elyes, Richards Stephen, Warren Wesley C., Waterhouse Robert M., Dillon Rod J., McDowell Mary Ann
Genomic Analysis of Two Phlebotomine Sand Fly Vectors of Leishmania from the New and Old World Journal Article
In: PLoS Negl. Trop. Dis., vol. 17, 2023.
Abstract | Links | BibTeX | Tags: Drosophila melanogaster, Genome annotation, Genomics, imler, Invertebrate genomics, Leishmania, M3i, Marques, mosquitoes, Phylogenetic analysis
@article{McDowell2023,
title = {Genomic Analysis of Two Phlebotomine Sand Fly Vectors of Leishmania from the New and Old World},
author = {Frédéric Labbé AND Maha Abdeladhim AND Jenica Abrudan AND Alejandra Saori Araki AND Ricardo N. Araujo AND Peter Arensburger AND Joshua B. Benoit AND Reginaldo Pecanha Brazil AND Rafaela V. Bruno AND Gustavo Bueno da Silva Rivas AND Vinicius Carvalho de Abreu AND Jason Charamis AND Iliano V. Coutinho-Abreu AND Samara G. da Costa-Latgé AND Alistair Darby AND Viv M. Dillon AND Scott J. Emrich AND Daniela Fernandez-Medina AND Nelder Figueiredo Gontijo AND Catherine M. Flanley AND Derek Gatherer AND Fernando A. Genta AND Sandra Gesing AND Gloria I. Giraldo-Calderón AND Bruno Gomes AND Eric Roberto Guimaraes Rocha Aguiar AND James G. C. Hamilton AND Omar Hamarsheh AND Mallory Hawksworth AND Jacob M. Hendershot AND Paul V. Hickner AND Jean-Luc Imler AND Panagiotis Ioannidis AND Emily C. Jennings AND Shaden Kamhawi AND Charikleia Karageorgiou AND Ryan C. Kennedy AND Andreas Krueger AND José M. Latorre-Estivalis AND Petros Ligoxygakis AND Antonio Carlos A. Meireles-Filho AND Patrick Minx AND Jose Carlos Miranda AND Michael J. Montague AND Ronald J. Nowling AND Fabiano Oliveira AND João Ortigão-Farias AND Marcio G. Pavan AND Marcos Horacio Pereira AND Andre Nobrega Pitaluga AND Roenick Proveti Olmo AND Marcelo Ramalho-Ortigao AND José M. C. Ribeiro AND Andrew J. Rosendale AND Mauricio R. V. Sant’Anna AND Steven E. Scherer AND Nágila F. C. Secundino AND Douglas A. Shoue AND Caroline da Silva Moraes AND João Silveira Moledo Gesto AND Nataly Araujo Souza AND Zainulabueddin Syed AND Samuel Tadros AND Rayane Teles-de-Freitas AND Erich L. Telleria AND Chad Tomlinson AND Yara M. Traub-Csekö AND João Trindade Marques AND Zhijian Tu AND Maria F. Unger AND Jesus Valenzuela AND Flávia V. Ferreira AND Karla P. V. de Oliveira AND Felipe M. Vigoder AND John Vontas AND Lihui Wang AND Gareth D. Weedall AND Elyes Zhioua AND Stephen Richards AND Wesley C. Warren AND Robert M. Waterhouse AND Rod J. Dillon AND Mary Ann McDowell},
url = {https://journals.plos.org/plosntds/article?id=10.1371/journal.pntd.0010862},
doi = {10.1371/journal.pntd.0010862},
year = {2023},
date = {2023-04-12},
urldate = {2023-04-12},
journal = {PLoS Negl. Trop. Dis.},
volume = {17},
abstract = {The leishmaniases are a group of neglected tropical diseases caused by protist parasites from the Genus Leishmania. Different Leishmania species present a wide clinical profile, ranging from mild, often self-resolving cutaneous lesions that can lead to protective immunity, to severe metastatic mucosal disease, to visceral disease that is ultimately fatal. Leishmania parasites are transmitted by the bites of sand flies, and as no approved human vaccine exists, available drugs are toxic and/or expensive and parasite resistance to them is emerging, new dual control strategies to combat these diseases must be developed, combining interventions on human infections and integrated sand fly population management. Effective vector control requires a comprehensive understanding of the biology of sand flies. To this end, we sequenced and annotated the genomes of two sand fly species that are important leishmaniasis vectors from the Old and New Worlds. These genomes allow us to better understand, at the genetic level, processes important in the vector biology of these species, such as finding hosts, blood-feeding, immunity, and detoxification. These genomic resources highlight the driving forces of evolution of two major Leishmania vectors and provide foundations for future research on how to better prevent leishmaniasis by control of the sand fly vectors.},
keywords = {Drosophila melanogaster, Genome annotation, Genomics, imler, Invertebrate genomics, Leishmania, M3i, Marques, mosquitoes, Phylogenetic analysis},
pubstate = {published},
tppubtype = {article}
}
The leishmaniases are a group of neglected tropical diseases caused by protist parasites from the Genus Leishmania. Different Leishmania species present a wide clinical profile, ranging from mild, often self-resolving cutaneous lesions that can lead to protective immunity, to severe metastatic mucosal disease, to visceral disease that is ultimately fatal. Leishmania parasites are transmitted by the bites of sand flies, and as no approved human vaccine exists, available drugs are toxic and/or expensive and parasite resistance to them is emerging, new dual control strategies to combat these diseases must be developed, combining interventions on human infections and integrated sand fly population management. Effective vector control requires a comprehensive understanding of the biology of sand flies. To this end, we sequenced and annotated the genomes of two sand fly species that are important leishmaniasis vectors from the Old and New Worlds. These genomes allow us to better understand, at the genetic level, processes important in the vector biology of these species, such as finding hosts, blood-feeding, immunity, and detoxification. These genomic resources highlight the driving forces of evolution of two major Leishmania vectors and provide foundations for future research on how to better prevent leishmaniasis by control of the sand fly vectors.
de Faria Isaque J. S., Imler Jean-Luc, Marques João T.
Protocol for the analysis of double-stranded RNAs in virus-infected insect cells using anti-dsRNA antibodies Journal Article
In: STAR Protocols, vol. 4, iss. 1, 2023.
Abstract | Links | BibTeX | Tags: antibody, cell bioloby, imler, Immunology, M3i, Marques, microbiology, Microscopy, Molecular Biology
@article{deFaria2023,
title = {Protocol for the analysis of double-stranded RNAs in virus-infected insect cells using anti-dsRNA antibodies},
author = {Isaque J.S. de Faria and Jean-Luc Imler and João T. Marques},
url = {https://doi.org/10.1016/j.xpro.2022.102033},
doi = {10.1016/j.xpro.2022.102033},
year = {2023},
date = {2023-03-17},
urldate = {2023-03-17},
journal = {STAR Protocols},
volume = {4},
issue = {1},
abstract = {Characterization of double-stranded (ds)RNAs is relevant to the understanding of viral replication and immune sensing. Here, we provide a protocol describing the use of anti-dsRNA antibodies for immunofluorescence and immunoblotting in virus-infected insect cells, which can also be applied to tissues and other organisms. We describe the procedures to prepare insect cells for viral infection, followed by RNA extraction and in vitro production of synthetic dsRNA controls. We then detail the steps for dsRNA detection by immunoblotting and immunofluorescence. For complete details on the use and execution of this protocol, please refer to de Faria et al. (2022).1},
keywords = {antibody, cell bioloby, imler, Immunology, M3i, Marques, microbiology, Microscopy, Molecular Biology},
pubstate = {published},
tppubtype = {article}
}
Characterization of double-stranded (ds)RNAs is relevant to the understanding of viral replication and immune sensing. Here, we provide a protocol describing the use of anti-dsRNA antibodies for immunofluorescence and immunoblotting in virus-infected insect cells, which can also be applied to tissues and other organisms. We describe the procedures to prepare insect cells for viral infection, followed by RNA extraction and in vitro production of synthetic dsRNA controls. We then detail the steps for dsRNA detection by immunoblotting and immunofluorescence. For complete details on the use and execution of this protocol, please refer to de Faria et al. (2022).1
Huang Jianqiong, Lou Yanyan, Liu Jiyong, Bulet Philippe, Cai Chuping, Ma Kaiyu, Jiao Renjie, Hoffmann Jules A, Liégeois Samuel, Lia Zi, Ferrandon Dominique
A Toll pathway effector protects Drosophila specifically from distinct toxins secreted by a fungus or a bacterium Journal Article
In: PNAS, vol. 120, no. 12, 2023.
Abstract | Links | BibTeX | Tags: Baramicin A, Destruxin A, disease tolerance, enterocin O16, ferrandon, hoffmann, M3i, microbial toxins, resilience
@article{Huang2023,
title = {A Toll pathway effector protects Drosophila specifically from distinct toxins secreted by a fungus or a bacterium},
author = {Jianqiong Huang and Yanyan Lou and Jiyong Liu and Philippe Bulet and Chuping Cai and Kaiyu Ma and Renjie Jiao and Jules A Hoffmann and Samuel Liégeois and Zi Lia and Dominique Ferrandon},
editor = {Hugo Bellen, Baylor College of Medicine, Houston, TX},
url = {https://doi.org/10.1073/pnas.2205140120},
doi = {10.1073/pnas.2205140120},
year = {2023},
date = {2023-03-14},
urldate = {2023-03-14},
journal = {PNAS},
volume = {120},
number = {12},
abstract = {Major immune response pathways control the expression of hundreds of genes that represent potential effectors of the immune response. The Drosophila Toll pathway is required in the host defenses against several Gram-positive bacterial infections as well as against fungal infections. The current paradigm is that peptides secreted in the hemolymph during the systemic immune response are either bona fide antimicrobial peptides or likely ones. The finding of a dual role for one Toll pathway effector in the resilience to both Enterococcus faecalis and Metarhizium robertsii infections underscores an original concept in insect innate immunity. Evolution can select effectors tailored to protect the host from the action of microbial toxins of prokaryotic or eukaryotic origin, independently of antibodies or detoxification pathways.},
keywords = {Baramicin A, Destruxin A, disease tolerance, enterocin O16, ferrandon, hoffmann, M3i, microbial toxins, resilience},
pubstate = {published},
tppubtype = {article}
}
Major immune response pathways control the expression of hundreds of genes that represent potential effectors of the immune response. The Drosophila Toll pathway is required in the host defenses against several Gram-positive bacterial infections as well as against fungal infections. The current paradigm is that peptides secreted in the hemolymph during the systemic immune response are either bona fide antimicrobial peptides or likely ones. The finding of a dual role for one Toll pathway effector in the resilience to both Enterococcus faecalis and Metarhizium robertsii infections underscores an original concept in insect innate immunity. Evolution can select effectors tailored to protect the host from the action of microbial toxins of prokaryotic or eukaryotic origin, independently of antibodies or detoxification pathways.
Klug Dennis, Gautier Amandine, Calvo Eric, Marois Eric, Blandin Stéphanie A.
The salivary protein Saglin facilitates efficient midgut colonization of Anopheles mosquitoes by malaria parasites Journal Article
In: Plos Pathogens, vol. 19, iss. 3, no. 3, 2023.
Abstract | Links | BibTeX | Tags: blandin, BLOOD, M3i, malarial parasites, marois, mosquitoes, Oocysts, Parasitic Diseases, Plasmodium, salivary glands, sporozoites
@article{Klug2023,
title = {The salivary protein Saglin facilitates efficient midgut colonization of Anopheles mosquitoes by malaria parasites},
author = {Dennis Klug and Amandine Gautier and Eric Calvo and Eric Marois and Stéphanie A. Blandin},
url = {https://doi.org/10.1371/journal.ppat.1010538},
doi = {10.1371/journal.ppat.1010538},
year = {2023},
date = {2023-03-02},
urldate = {2023-03-02},
booktitle = {Plos Pathogens},
journal = {Plos Pathogens},
volume = {19},
number = {3},
issue = {3},
abstract = {Female mosquitoes rely on blood feeding to acquire sufficient nutrients for egg development. Because of the importance of this process mosquitoes evolved salivary proteins with a broad range of functions acting as blood thinners, anti-coagulants and immunosuppressants. The effect of these proteins on the blood at the bite site directly influences the size of the blood bolus a female takes up in a given time frame. Both, time of feeding and bolus size, are important parameters for fecundity and survival. Recent studies have shown that a significant proportion of salivated proteins is re-ingested during feeding and becomes part of the blood meal. Here we investigated the salivary protein Saglin which has been previously suggested as putative receptor mediating malaria parasite entry into the salivary gland. By engineering a loss-of-function mutant in An. coluzzi we could show that the absence of Saglin impairs the development of parasite stages in the blood meal of the rodent malaria parasite P. berghei and the human malaria parasite P. falciparum lowering the parasite burden of subsequent stages and preventing efficient transmission at low infection densities. Furthermore, we could show that Saglin is present in the blood meal after feeding possibly indicating a previously overlooked parasite-vector interaction.},
keywords = {blandin, BLOOD, M3i, malarial parasites, marois, mosquitoes, Oocysts, Parasitic Diseases, Plasmodium, salivary glands, sporozoites},
pubstate = {published},
tppubtype = {article}
}
Female mosquitoes rely on blood feeding to acquire sufficient nutrients for egg development. Because of the importance of this process mosquitoes evolved salivary proteins with a broad range of functions acting as blood thinners, anti-coagulants and immunosuppressants. The effect of these proteins on the blood at the bite site directly influences the size of the blood bolus a female takes up in a given time frame. Both, time of feeding and bolus size, are important parameters for fecundity and survival. Recent studies have shown that a significant proportion of salivated proteins is re-ingested during feeding and becomes part of the blood meal. Here we investigated the salivary protein Saglin which has been previously suggested as putative receptor mediating malaria parasite entry into the salivary gland. By engineering a loss-of-function mutant in An. coluzzi we could show that the absence of Saglin impairs the development of parasite stages in the blood meal of the rodent malaria parasite P. berghei and the human malaria parasite P. falciparum lowering the parasite burden of subsequent stages and preventing efficient transmission at low infection densities. Furthermore, we could show that Saglin is present in the blood meal after feeding possibly indicating a previously overlooked parasite-vector interaction.
R.P. Olmo, Y.M.H. Todjro, E.R.G.R. Aguiar, de Almeida J.P.P., F.V. Ferreira, J.N. Armache, de Faria I.J.S., A.G.A. Ferreira, S.C.D. Amadou, A.T.S. Silva, de Souza K.P.R., A.P.P. Vilela, A. Babarit, C.H. Tan, M. Diallo, A. Gaye, C. Paupy, J. Obame-Nkoghe, T.M. Visser, C.J.M. Koenraadt, M.A. Wongsokarijo, A.L.C. Cruz, M.T. Prieto, M.C.P. Parra, Nogueira M.L., V. Avelino-Silva, R.N. Mota, M.A.Z. Borges, B.P. Drumond, E.G. Kroon, M. Recker, L. Sedda, E. Marois, J.L. Imler, J.T. Marques
Mosquito vector competence for dengue is modulated by insect-specific viruses Journal Article
In: Nature Microbiology, vol. 8, iss. 1, 2023.
Abstract | Links | BibTeX | Tags: Dengue, imler, M3i, marois, Marques, mosquito, Olmo, virus
@article{Olmo.2023,
title = {Mosquito vector competence for dengue is modulated by insect-specific viruses},
author = {Olmo R.P. and Todjro Y.M.H. and Aguiar E.R.G.R. and de Almeida J.P.P. and Ferreira F.V. and Armache J.N. and de Faria I.J.S. and Ferreira A.G.A. and Amadou S.C.D. and Silva A.T.S. and de Souza K.P.R. and Vilela A.P.P. and Babarit A. and Tan C.H. and Diallo M. and Gaye A. and Paupy C. and Obame-Nkoghe J. and Visser T.M. and Koenraadt C.J.M. and Wongsokarijo M.A. and Cruz A.L.C. and Prieto M.T. and Parra M.C.P. and Nogueira M.L., and Avelino-Silva V. and Mota R.N. and Borges M.A.Z. and Drumond B.P. and Kroon E.G. and Recker M. and Sedda L. and Marois E. and Imler J.L. and Marques J.T. },
url = {https://doi.org/},
doi = {10.1038/s41564-022-01289-4},
year = {2023},
date = {2023-01-05},
urldate = {2023-01-05},
journal = {Nature Microbiology},
volume = {8},
issue = {1},
abstract = {Aedes aegypti and A. albopictus mosquitoes are the main vectors for dengue virus (DENV) and other arboviruses, including Zika virus (ZIKV). Understanding the factors that affect transmission of arboviruses from mosquitoes to humans is a priority because it could inform public health and targeted interventions. Reasoning that interactions among viruses in the vector insect might affect transmission, we analysed the viromes of 815 urban Aedes mosquitoes collected from 12 countries worldwide. Two mosquito-specific viruses, Phasi Charoen-like virus (PCLV) and Humaita Tubiacanga virus (HTV), were the most abundant in A. aegypti worldwide. Spatiotemporal analyses of virus circulation in an endemic urban area revealed a 200% increase in chances of having DENV in wild A. aegypti mosquitoes when both HTV and PCLV were present. Using a mouse model in the laboratory, we showed that the presence of HTV and PCLV increased the ability of mosquitoes to transmit DENV and ZIKV to a vertebrate host. By transcriptomic analysis, we found that in DENV-infected mosquitoes, HTV and PCLV block the downregulation of histone H4, which we identify as an important proviral host factor in vivo.},
keywords = {Dengue, imler, M3i, marois, Marques, mosquito, Olmo, virus},
pubstate = {published},
tppubtype = {article}
}
Aedes aegypti and A. albopictus mosquitoes are the main vectors for dengue virus (DENV) and other arboviruses, including Zika virus (ZIKV). Understanding the factors that affect transmission of arboviruses from mosquitoes to humans is a priority because it could inform public health and targeted interventions. Reasoning that interactions among viruses in the vector insect might affect transmission, we analysed the viromes of 815 urban Aedes mosquitoes collected from 12 countries worldwide. Two mosquito-specific viruses, Phasi Charoen-like virus (PCLV) and Humaita Tubiacanga virus (HTV), were the most abundant in A. aegypti worldwide. Spatiotemporal analyses of virus circulation in an endemic urban area revealed a 200% increase in chances of having DENV in wild A. aegypti mosquitoes when both HTV and PCLV were present. Using a mouse model in the laboratory, we showed that the presence of HTV and PCLV increased the ability of mosquitoes to transmit DENV and ZIKV to a vertebrate host. By transcriptomic analysis, we found that in DENV-infected mosquitoes, HTV and PCLV block the downregulation of histone H4, which we identify as an important proviral host factor in vivo.
2022
Xu Rui, Lou Yanyan, Tidu Antonin, Bulet Philippe, Heinekamp Thorsten, Martin Franck, Brakhage Axel, Li Zi, Liégeois Samuel, Ferrandon Dominique
The Toll pathway mediates Drosophila resilience to Aspergillus mycotoxins through specific Bomanins Journal Article
In: EMBO Rep, pp. e56036, 2022, ISSN: 1469-3178.
Abstract | Links | BibTeX | Tags: ERIANI, ferrandon, M3i, MARTIN, Unité ARN
@article{pmid36322050,
title = {The Toll pathway mediates Drosophila resilience to Aspergillus mycotoxins through specific Bomanins},
author = {Rui Xu and Yanyan Lou and Antonin Tidu and Philippe Bulet and Thorsten Heinekamp and Franck Martin and Axel Brakhage and Zi Li and Samuel Liégeois and Dominique Ferrandon},
url = {https://pubmed.ncbi.nlm.nih.gov/36322050/},
doi = {10.15252/embr.202256036},
issn = {1469-3178},
year = {2022},
date = {2022-11-01},
urldate = {2022-11-01},
journal = {EMBO Rep},
pages = {e56036},
abstract = {Host defense against infections encompasses both resistance, which targets microorganisms for neutralization or elimination, and resilience/disease tolerance, which allows the host to withstand/tolerate pathogens and repair damages. In Drosophila, the Toll signaling pathway is thought to mediate resistance against fungal infections by regulating the secretion of antimicrobial peptides, potentially including Bomanins. We find that Aspergillus fumigatus kills Drosophila Toll pathway mutants without invasion because its dissemination is blocked by melanization, suggesting a role for Toll in host defense distinct from resistance. We report that mutants affecting the Toll pathway or the 55C Bomanin locus are susceptible to the injection of two Aspergillus mycotoxins, restrictocin and verruculogen. The vulnerability of 55C deletion mutants to these mycotoxins is rescued by the overexpression of Bomanins specific to each challenge. Mechanistically, flies in which BomS6 is expressed in the nervous system exhibit an enhanced recovery from the tremors induced by injected verruculogen and display improved survival. Thus, innate immunity also protects the host against the action of microbial toxins through secreted peptides and thereby increases its resilience to infection.},
keywords = {ERIANI, ferrandon, M3i, MARTIN, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Host defense against infections encompasses both resistance, which targets microorganisms for neutralization or elimination, and resilience/disease tolerance, which allows the host to withstand/tolerate pathogens and repair damages. In Drosophila, the Toll signaling pathway is thought to mediate resistance against fungal infections by regulating the secretion of antimicrobial peptides, potentially including Bomanins. We find that Aspergillus fumigatus kills Drosophila Toll pathway mutants without invasion because its dissemination is blocked by melanization, suggesting a role for Toll in host defense distinct from resistance. We report that mutants affecting the Toll pathway or the 55C Bomanin locus are susceptible to the injection of two Aspergillus mycotoxins, restrictocin and verruculogen. The vulnerability of 55C deletion mutants to these mycotoxins is rescued by the overexpression of Bomanins specific to each challenge. Mechanistically, flies in which BomS6 is expressed in the nervous system exhibit an enhanced recovery from the tremors induced by injected verruculogen and display improved survival. Thus, innate immunity also protects the host against the action of microbial toxins through secreted peptides and thereby increases its resilience to infection.
Cammarata-Mouchtouris Alexandre, Acker Adrian, Goto Akira, Chen Di, Matt Nicolas, Leclerc Vincent
Dynamic Regulation of NF-κB Response in Innate Immunity: The Case of the IMD Pathway in Drosophila Journal Article
In: Biomedicines, vol. 10, no. 9, 2022, ISSN: 2227-9059.
Abstract | Links | BibTeX | Tags: M3i, matt
@article{pmid36140409,
title = {Dynamic Regulation of NF-κB Response in Innate Immunity: The Case of the IMD Pathway in Drosophila},
author = {Alexandre Cammarata-Mouchtouris and Adrian Acker and Akira Goto and Di Chen and Nicolas Matt and Vincent Leclerc},
doi = {10.3390/biomedicines10092304},
issn = {2227-9059},
year = {2022},
date = {2022-09-01},
urldate = {2022-09-01},
journal = {Biomedicines},
volume = {10},
number = {9},
abstract = {Metazoans have developed strategies to protect themselves from pathogenic attack. These preserved mechanisms constitute the immune system, composed of innate and adaptive responses. Among the two kinds, the innate immune system involves the activation of a fast response. NF-κB signaling pathways are activated during infections and lead to the expression of timely-controlled immune response genes. However, activation of NF-κB pathways can be deleterious when uncontrolled. Their regulation is necessary to prevent the development of inflammatory diseases or cancers. The similarity of the NF-κB pathways mediating immune mechanisms in insects and mammals makes a suitable model for studying the innate immune response and learning general mechanisms that are also relevant for humans. In this review, we summarize what is known about the dynamic regulation of the central NF-κB-pathways and go into detail on the molecular level of the IMD pathway. We report on the role of the nuclear protein Akirin in the regulation of the NF-κB Relish immune response. The use of the model allows the understanding of the fine-tuned regulation of this central NF-κB pathway.},
keywords = {M3i, matt},
pubstate = {published},
tppubtype = {article}
}
Metazoans have developed strategies to protect themselves from pathogenic attack. These preserved mechanisms constitute the immune system, composed of innate and adaptive responses. Among the two kinds, the innate immune system involves the activation of a fast response. NF-κB signaling pathways are activated during infections and lead to the expression of timely-controlled immune response genes. However, activation of NF-κB pathways can be deleterious when uncontrolled. Their regulation is necessary to prevent the development of inflammatory diseases or cancers. The similarity of the NF-κB pathways mediating immune mechanisms in insects and mammals makes a suitable model for studying the innate immune response and learning general mechanisms that are also relevant for humans. In this review, we summarize what is known about the dynamic regulation of the central NF-κB-pathways and go into detail on the molecular level of the IMD pathway. We report on the role of the nuclear protein Akirin in the regulation of the NF-κB Relish immune response. The use of the model allows the understanding of the fine-tuned regulation of this central NF-κB pathway.
de Faria Isaque J. S., Aguiar Eric R. G. R., Olmo Roenick P., da Silva Juliana Alves, Daeffler Laurent, Carthew Richard W., Imler Jean-Luc, Marques Joao T.
Invading viral DNA triggers dsRNA synthesis by RNA polymerase II to activate antiviral RNA interference in Drosophila Journal Article
In: Cell Reports, vol. 39, pp. 110976, 2022.
Abstract | Links | BibTeX | Tags: antiviral, Drosophila, dsRNA, imler, M3i, Marques, protocol, RNA Interference
@article{dedaMarques2022,
title = {Invading viral DNA triggers dsRNA synthesis by RNA polymerase II to activate antiviral RNA interference in Drosophila},
author = {Isaque J.S. de Faria and Eric R.G.R. Aguiar and Roenick P. Olmo and Juliana Alves da Silva and Laurent Daeffler and Richard W. Carthew and Jean-Luc Imler and Joao T. Marques},
doi = {10.1016/j.celrep.2022.110976},
year = {2022},
date = {2022-06-21},
urldate = {2022-06-21},
journal = {Cell Reports},
volume = {39},
pages = {110976},
abstract = {dsRNA sensing triggers antiviral responses against RNA and DNA viruses in diverse eukaryotes. In Drosophila, Invertebrate iridescent virus 6 (IIV-6), a large DNA virus, triggers production of small interfering RNAs (siRNAs) by the dsRNA sensor Dicer-2. Here, we show that host RNA polymerase II (RNAPII) bidirec- tionally transcribes specific AT-rich regions of the IIV-6 DNA genome to generate dsRNA. Both replicative and naked IIV-6 genomes trigger production of dsRNA in Drosophila cells, implying direct sensing of invading DNA. Loquacious-PD, a Dicer-2 co-factor essential for the biogenesis of endogenous siRNAs, is dispensable for processing of IIV-6-derived dsRNAs, which suggests that they are distinct. Consistent with this finding, inhibition of the RNAPII co-factor P-TEFb affects the synthesis of endogenous, but not virus-derived, dsRNA. Altogether, our results suggest that a non-canonical RNAPII complex recognizes invading viral DNA to synthesize virus-derived dsRNA, which activates the antiviral siRNA pathway in Drosophila.},
keywords = {antiviral, Drosophila, dsRNA, imler, M3i, Marques, protocol, RNA Interference},
pubstate = {published},
tppubtype = {article}
}
dsRNA sensing triggers antiviral responses against RNA and DNA viruses in diverse eukaryotes. In Drosophila, Invertebrate iridescent virus 6 (IIV-6), a large DNA virus, triggers production of small interfering RNAs (siRNAs) by the dsRNA sensor Dicer-2. Here, we show that host RNA polymerase II (RNAPII) bidirec- tionally transcribes specific AT-rich regions of the IIV-6 DNA genome to generate dsRNA. Both replicative and naked IIV-6 genomes trigger production of dsRNA in Drosophila cells, implying direct sensing of invading DNA. Loquacious-PD, a Dicer-2 co-factor essential for the biogenesis of endogenous siRNAs, is dispensable for processing of IIV-6-derived dsRNAs, which suggests that they are distinct. Consistent with this finding, inhibition of the RNAPII co-factor P-TEFb affects the synthesis of endogenous, but not virus-derived, dsRNA. Altogether, our results suggest that a non-canonical RNAPII complex recognizes invading viral DNA to synthesize virus-derived dsRNA, which activates the antiviral siRNA pathway in Drosophila.
Cai H, Meignin C, Imler JL
cGAS-like receptor-mediated immunity: the insect perspective Journal Article
In: Current Opinion in Immunology, vol. 74, pp. 183-189, 2022.
Abstract | Links | BibTeX | Tags: imler, M3i, meignin
@article{Imler2022,
title = {cGAS-like receptor-mediated immunity: the insect perspective},
author = {H Cai and C Meignin and JL Imler},
doi = {10.1016/j.coi.2022.01.005},
year = {2022},
date = {2022-02-08},
urldate = {2022-02-08},
journal = {Current Opinion in Immunology},
volume = {74},
pages = {183-189},
abstract = {The cGAS-STING pathway plays a central role in the detection of DNA in the cytosol of mammalian cells and activation of immunity. Although the early evolutionary origin of this pathway in animals has been noted, its ancestral functions have remained elusive so far. We review here new findings in invertebrates establishing a role in sensing and signaling infection, triggering potent transcriptional responses, in addition to autophagy. Results from flies and moths/butterflies points to the importance of STING signaling in antiviral immunity in insects. The recent characterization of cGAS-like receptors in Drosophila reveals the plasticity of this family of pattern-recognition receptors, able to accommodate ligands different from DNA and to produce cyclic dinucleotides beyond 2′3′-cGAMP.},
keywords = {imler, M3i, meignin},
pubstate = {published},
tppubtype = {article}
}
The cGAS-STING pathway plays a central role in the detection of DNA in the cytosol of mammalian cells and activation of immunity. Although the early evolutionary origin of this pathway in animals has been noted, its ancestral functions have remained elusive so far. We review here new findings in invertebrates establishing a role in sensing and signaling infection, triggering potent transcriptional responses, in addition to autophagy. Results from flies and moths/butterflies points to the importance of STING signaling in antiviral immunity in insects. The recent characterization of cGAS-like receptors in Drosophila reveals the plasticity of this family of pattern-recognition receptors, able to accommodate ligands different from DNA and to produce cyclic dinucleotides beyond 2′3′-cGAMP.
2021
Pennemann Friederike L, Mussabekova Assel, Urban Christian, Stukalov Alexey, Andersen Line Lykke, Grass Vincent, Lavacca Teresa Maria, Holze Cathleen, Oubraham Lila, Benamrouche Yasmine, Girardi Enrico, Boulos Rasha E, Hartmann Rune, Superti-Furga Giulio, Habjan Matthias, Imler Jean-Luc, Meignin Carine, Pichlmair Andreas
Cross-species analysis of viral nucleic acid interacting proteins identifies TAOKs as innate immune regulators Journal Article
In: Nat Commun, vol. 12, no. 1, pp. 7009, 2021, ISSN: 2041-1723.
Abstract | Links | BibTeX | Tags: antiviral innate immunity, imler, M3i, meignin
@article{pmid34853303,
title = {Cross-species analysis of viral nucleic acid interacting proteins identifies TAOKs as innate immune regulators},
author = {Friederike L Pennemann and Assel Mussabekova and Christian Urban and Alexey Stukalov and Line Lykke Andersen and Vincent Grass and Teresa Maria Lavacca and Cathleen Holze and Lila Oubraham and Yasmine Benamrouche and Enrico Girardi and Rasha E Boulos and Rune Hartmann and Giulio Superti-Furga and Matthias Habjan and Jean-Luc Imler and Carine Meignin and Andreas Pichlmair},
doi = {10.1038/s41467-021-27192-w},
issn = {2041-1723},
year = {2021},
date = {2021-12-01},
urldate = {2021-12-01},
journal = {Nat Commun},
volume = {12},
number = {1},
pages = {7009},
abstract = {The cell intrinsic antiviral response of multicellular organisms developed over millions of years and critically relies on the ability to sense and eliminate viral nucleic acids. Here we use an affinity proteomics approach in evolutionary distant species (human, mouse and fly) to identify proteins that are conserved in their ability to associate with diverse viral nucleic acids. This approach shows a core of orthologous proteins targeting viral genetic material and species-specific interactions. Functional characterization of the influence of 181 candidates on replication of 6 distinct viruses in human cells and flies identifies 128 nucleic acid binding proteins with an impact on virus growth. We identify the family of TAO kinases (TAOK1, -2 and -3) as dsRNA-interacting antiviral proteins and show their requirement for type-I interferon induction. Depletion of TAO kinases in mammals or flies leads to an impaired response to virus infection characterized by a reduced induction of interferon stimulated genes in mammals and impaired expression of srg1 and diedel in flies. Overall, our study shows a larger set of proteins able to mediate the interaction between viral genetic material and host factors than anticipated so far, attesting to the ancestral roots of innate immunity and to the lineage-specific pressures exerted by viruses.},
keywords = {antiviral innate immunity, imler, M3i, meignin},
pubstate = {published},
tppubtype = {article}
}
The cell intrinsic antiviral response of multicellular organisms developed over millions of years and critically relies on the ability to sense and eliminate viral nucleic acids. Here we use an affinity proteomics approach in evolutionary distant species (human, mouse and fly) to identify proteins that are conserved in their ability to associate with diverse viral nucleic acids. This approach shows a core of orthologous proteins targeting viral genetic material and species-specific interactions. Functional characterization of the influence of 181 candidates on replication of 6 distinct viruses in human cells and flies identifies 128 nucleic acid binding proteins with an impact on virus growth. We identify the family of TAO kinases (TAOK1, -2 and -3) as dsRNA-interacting antiviral proteins and show their requirement for type-I interferon induction. Depletion of TAO kinases in mammals or flies leads to an impaired response to virus infection characterized by a reduced induction of interferon stimulated genes in mammals and impaired expression of srg1 and diedel in flies. Overall, our study shows a larger set of proteins able to mediate the interaction between viral genetic material and host factors than anticipated so far, attesting to the ancestral roots of innate immunity and to the lineage-specific pressures exerted by viruses.
Wehbe Layale Salem, Barakat Dana, Acker Adrian, Khoury Rita El, Reichhart Jean-Marc, Matt Nicolas, Chamy Laure El
Protein Phosphatase 4 Negatively Regulates the Immune Deficiency-NF-κB Pathway during the Immune Response Journal Article
In: J Immunol, vol. 207, no. 6, pp. 1616–1626, 2021, ISSN: 1550-6606.
Abstract | Links | BibTeX | Tags: Drosophila, IKK complex, IMD, immune response, M3i, matt, NF-κB, PP4 complex
@article{pmid34452932,
title = {Protein Phosphatase 4 Negatively Regulates the Immune Deficiency-NF-κB Pathway during the Immune Response},
author = {Layale Salem Wehbe and Dana Barakat and Adrian Acker and Rita El Khoury and Jean-Marc Reichhart and Nicolas Matt and Laure El Chamy},
doi = {10.4049/jimmunol.1901497},
issn = {1550-6606},
year = {2021},
date = {2021-08-27},
urldate = {2021-08-27},
journal = {J Immunol},
volume = {207},
number = {6},
pages = {1616--1626},
abstract = {The evolutionarily conserved immune deficiency (IMD) signaling pathway shields against bacterial infections. It regulates the expression of antimicrobial peptides encoding genes through the activation of the NF-κB transcription factor Relish. Tight regulation of the signaling cascade ensures a balanced immune response, which is otherwise highly harmful. Several phosphorylation events mediate intracellular progression of the IMD pathway. However, signal termination by dephosphorylation remains largely elusive. Here, we identify the highly conserved protein phosphatase 4 (PP4) complex as a bona fide negative regulator of the IMD pathway. RNA interference-mediated gene silencing of , , and which encode the catalytic and regulatory subunits of the phosphatase complex, respectively, caused a marked upregulation of bacterial-induced antimicrobial peptide gene expression in both S2 cells and adult flies. Deregulated IMD signaling is associated with reduced lifespan of -deficient flies in the absence of any infection. In contrast, flies overexpressing this phosphatase are highly sensitive to bacterial infections. Altogether, our results highlight an evolutionarily conserved function of PP4c in the regulation of NF-κB signaling from to mammals.},
keywords = {Drosophila, IKK complex, IMD, immune response, M3i, matt, NF-κB, PP4 complex},
pubstate = {published},
tppubtype = {article}
}
The evolutionarily conserved immune deficiency (IMD) signaling pathway shields against bacterial infections. It regulates the expression of antimicrobial peptides encoding genes through the activation of the NF-κB transcription factor Relish. Tight regulation of the signaling cascade ensures a balanced immune response, which is otherwise highly harmful. Several phosphorylation events mediate intracellular progression of the IMD pathway. However, signal termination by dephosphorylation remains largely elusive. Here, we identify the highly conserved protein phosphatase 4 (PP4) complex as a bona fide negative regulator of the IMD pathway. RNA interference-mediated gene silencing of , , and which encode the catalytic and regulatory subunits of the phosphatase complex, respectively, caused a marked upregulation of bacterial-induced antimicrobial peptide gene expression in both S2 cells and adult flies. Deregulated IMD signaling is associated with reduced lifespan of -deficient flies in the absence of any infection. In contrast, flies overexpressing this phosphatase are highly sensitive to bacterial infections. Altogether, our results highlight an evolutionarily conserved function of PP4c in the regulation of NF-κB signaling from to mammals.
Loiseau Vincent, Peccoud Jean, Bouzar Clémence, Guillier Sandra, Fan Jiangbin, Alletti Gianpiero Gueli, Meignin Carine, Herniou Elisabeth A, Federici Brian A, Wennmann Jörg T, Jehle Johannes A, Cordaux Richard, Gilbert Clément
Monitoring Insect Transposable Elements in Large Double-Stranded DNA Viruses Reveals Host-to-Virus and Virus-to-Virus Transposition Journal Article
In: Mol Biol Evol, vol. 38, no. 9, pp. 3512–3530, 2021, ISSN: 1537-1719.
Abstract | Links | BibTeX | Tags: M3i, meignin
@article{pmid34191026,
title = {Monitoring Insect Transposable Elements in Large Double-Stranded DNA Viruses Reveals Host-to-Virus and Virus-to-Virus Transposition},
author = {Vincent Loiseau and Jean Peccoud and Clémence Bouzar and Sandra Guillier and Jiangbin Fan and Gianpiero Gueli Alletti and Carine Meignin and Elisabeth A Herniou and Brian A Federici and Jörg T Wennmann and Johannes A Jehle and Richard Cordaux and Clément Gilbert},
doi = {10.1093/molbev/msab198},
issn = {1537-1719},
year = {2021},
date = {2021-08-01},
urldate = {2021-08-01},
journal = {Mol Biol Evol},
volume = {38},
number = {9},
pages = {3512--3530},
abstract = {The mechanisms by which transposable elements (TEs) can be horizontally transferred between animals are unknown, but viruses are possible candidate vectors. Here, we surveyed the presence of host-derived TEs in viral genomes in 35 deep sequencing data sets produced from 11 host-virus systems, encompassing nine arthropod host species (five lepidopterans, two dipterans, and two crustaceans) and six different double-stranded (ds) DNA viruses (four baculoviruses and two iridoviruses). We found evidence of viral-borne TEs in 14 data sets, with frequencies of viral genomes carrying a TE ranging from 0.01% to 26.33% for baculoviruses and from 0.45% to 7.36% for iridoviruses. The analysis of viral populations separated by a single replication cycle revealed that viral-borne TEs originating from an initial host species can be retrieved after viral replication in another host species, sometimes at higher frequencies. Furthermore, we detected a strong increase in the number of integrations in a viral population for a TE absent from the hosts' genomes, indicating that this TE has undergone intense transposition within the viral population. Finally, we provide evidence that many TEs found integrated in viral genomes (15/41) have been horizontally transferred in insects. Altogether, our results indicate that multiple large dsDNA viruses have the capacity to shuttle TEs in insects and they underline the potential of viruses to act as vectors of horizontal transfer of TEs. Furthermore, the finding that TEs can transpose between viral genomes of a viral species sets viruses as possible new niches in which TEs can persist and evolve.},
keywords = {M3i, meignin},
pubstate = {published},
tppubtype = {article}
}
The mechanisms by which transposable elements (TEs) can be horizontally transferred between animals are unknown, but viruses are possible candidate vectors. Here, we surveyed the presence of host-derived TEs in viral genomes in 35 deep sequencing data sets produced from 11 host-virus systems, encompassing nine arthropod host species (five lepidopterans, two dipterans, and two crustaceans) and six different double-stranded (ds) DNA viruses (four baculoviruses and two iridoviruses). We found evidence of viral-borne TEs in 14 data sets, with frequencies of viral genomes carrying a TE ranging from 0.01% to 26.33% for baculoviruses and from 0.45% to 7.36% for iridoviruses. The analysis of viral populations separated by a single replication cycle revealed that viral-borne TEs originating from an initial host species can be retrieved after viral replication in another host species, sometimes at higher frequencies. Furthermore, we detected a strong increase in the number of integrations in a viral population for a TE absent from the hosts' genomes, indicating that this TE has undergone intense transposition within the viral population. Finally, we provide evidence that many TEs found integrated in viral genomes (15/41) have been horizontally transferred in insects. Altogether, our results indicate that multiple large dsDNA viruses have the capacity to shuttle TEs in insects and they underline the potential of viruses to act as vectors of horizontal transfer of TEs. Furthermore, the finding that TEs can transpose between viral genomes of a viral species sets viruses as possible new niches in which TEs can persist and evolve.
Prakash Pragya, Roychowdhury-Sinha Arghyashree, Goto Akira
Verloren negatively regulates the expression of IMD pathway dependent antimicrobial peptides in Drosophila Journal Article
In: Scientific Reports, vol. 11, no. 15549, 2021.
Abstract | Links | BibTeX | Tags: bacteria, Biochemistry, DNA, Fungi, Gene Expression, gene regulation, Genetics, hoffmann, Immunochemistry, Immunology, infection, inflammation, Innate immune cells, innate immunity, M3i, microbiology, Molecular Biology, pathogens, RNA, RNAi, Signal Transduction, Transcription
@article{Goto2021,
title = {Verloren negatively regulates the expression of IMD pathway dependent antimicrobial peptides in Drosophila},
author = {Pragya Prakash and Arghyashree Roychowdhury-Sinha and Akira Goto},
url = {https://www.nature.com/articles/s41598-021-94973-0},
doi = {10.1038/s41598-021-94973-0},
year = {2021},
date = {2021-07-30},
journal = {Scientific Reports},
volume = {11},
number = {15549},
abstract = {Drosophila immune deficiency (IMD) pathway is similar to the human tumor necrosis factor receptor (TNFR) signaling pathway and is preferentially activated by Gram-negative bacterial infection. Recent studies highlighted the importance of IMD pathway regulation as it is tightly controlled by numbers of negative regulators at multiple levels. Here, we report a new negative regulator of the IMD pathway, Verloren (Velo). Silencing of Velo led to constitutive expression of the IMD pathway dependent antimicrobial peptides (AMPs), and Escherichia coli stimulation further enhanced the AMP expression. Epistatic analysis indicated that Velo knock-down mediated AMP upregulation is dependent on the canonical members of the IMD pathway. The immune fluorescent study using overexpression constructs revealed that Velo resides both in the nucleus and cytoplasm, but the majority (~ 75%) is localized in the nucleus. We also observed from in vivo analysis that Velo knock-down flies exhibit significant upregulation of the AMP expression and reduced bacterial load. Survival experiments showed that Velo knock-down flies have a short lifespan and are susceptible to the infection of pathogenic Gram-negative bacteria, P. aeruginosa. Taken together, these data suggest that Velo is an additional new negative regulator of the IMD pathway, possibly acting in both the nucleus and cytoplasm.},
keywords = {bacteria, Biochemistry, DNA, Fungi, Gene Expression, gene regulation, Genetics, hoffmann, Immunochemistry, Immunology, infection, inflammation, Innate immune cells, innate immunity, M3i, microbiology, Molecular Biology, pathogens, RNA, RNAi, Signal Transduction, Transcription},
pubstate = {published},
tppubtype = {article}
}
Drosophila immune deficiency (IMD) pathway is similar to the human tumor necrosis factor receptor (TNFR) signaling pathway and is preferentially activated by Gram-negative bacterial infection. Recent studies highlighted the importance of IMD pathway regulation as it is tightly controlled by numbers of negative regulators at multiple levels. Here, we report a new negative regulator of the IMD pathway, Verloren (Velo). Silencing of Velo led to constitutive expression of the IMD pathway dependent antimicrobial peptides (AMPs), and Escherichia coli stimulation further enhanced the AMP expression. Epistatic analysis indicated that Velo knock-down mediated AMP upregulation is dependent on the canonical members of the IMD pathway. The immune fluorescent study using overexpression constructs revealed that Velo resides both in the nucleus and cytoplasm, but the majority (~ 75%) is localized in the nucleus. We also observed from in vivo analysis that Velo knock-down flies exhibit significant upregulation of the AMP expression and reduced bacterial load. Survival experiments showed that Velo knock-down flies have a short lifespan and are susceptible to the infection of pathogenic Gram-negative bacteria, P. aeruginosa. Taken together, these data suggest that Velo is an additional new negative regulator of the IMD pathway, possibly acting in both the nucleus and cytoplasm.
Holleufer Andreas, Winther Kasper Grønbjerg, Gad Hans Henrik, Ai Xianlong, Chen Yuqiang, Li Lihua, Wei Ziming, Deng Huimin, Liu Jiyong, Frederiksen Ninna Ahlmann, Simonsen Bine, Andersen Line Lykke, Kleigrewe Karin, Dalskov Louise, Pichlmair Andreas, Cai Hua, Imler Jean-Luc, Hartmann Rune
Two cGAS-like receptors induce antiviral immunity in Drosophila Journal Article
In: Nature, vol. 597, pp. 114-118, 2021.
Abstract | Links | BibTeX | Tags: antiviral immunity, cGAS-like receptors, Drosophila, imler, M3i
@article{Hartmann2021,
title = {Two cGAS-like receptors induce antiviral immunity in Drosophila},
author = {Andreas Holleufer and Kasper Grønbjerg Winther and Hans Henrik Gad and Xianlong Ai and Yuqiang Chen and Lihua Li and Ziming Wei and Huimin Deng and Jiyong Liu and Ninna Ahlmann Frederiksen and Bine Simonsen and Line Lykke Andersen and Karin Kleigrewe and Louise Dalskov and Andreas Pichlmair and Hua Cai and Jean-Luc Imler and Rune Hartmann},
editor = {Nature Publishing Group},
doi = {https://doi.org/10.1038/s41586-021-03800-z},
year = {2021},
date = {2021-07-14},
urldate = {2021-07-14},
journal = {Nature},
volume = {597},
pages = {114-118},
abstract = {In mammals, cyclic GMP-AMP (cGAMP) synthase (cGAS) produces the cyclic dinucleotide (CDN) 2'3'-cGAMP in response to cytosolic DNA and this triggers an antiviral immune response. cGAS belongs to a large family of cGAS/DncV-like nucleotidyltransferases, present in both prokaryotes1 and eukaryotes2–5. In bacteria, these enzymes synthesize a range of cyclic oligonucleotide and have recently emerged as important regulators of phage infections6–8. Here, we identify two novel cGAS-like receptors (cGLRs) in the insect Drosophila melanogaster. We show that cGLR1 and cGLR2 activate Sting and NF-κB dependent antiviral immunity in response to infection with RNA or DNA viruses. cGLR1 is activated by dsRNA to produce the novel CDN 3'2'-cGAMP whereas cGLR2 produces a combination of 2'3'-cGAMP and 3'2' cGAMP in response to a yet unidentified stimulus. Our data establish cGAS as the founding member of a family of receptors sensing different types of nucleic acids and triggering immunity through production of CDNs beyond 2'3'-cGAMP.},
keywords = {antiviral immunity, cGAS-like receptors, Drosophila, imler, M3i},
pubstate = {published},
tppubtype = {article}
}
In mammals, cyclic GMP-AMP (cGAMP) synthase (cGAS) produces the cyclic dinucleotide (CDN) 2'3'-cGAMP in response to cytosolic DNA and this triggers an antiviral immune response. cGAS belongs to a large family of cGAS/DncV-like nucleotidyltransferases, present in both prokaryotes1 and eukaryotes2–5. In bacteria, these enzymes synthesize a range of cyclic oligonucleotide and have recently emerged as important regulators of phage infections6–8. Here, we identify two novel cGAS-like receptors (cGLRs) in the insect Drosophila melanogaster. We show that cGLR1 and cGLR2 activate Sting and NF-κB dependent antiviral immunity in response to infection with RNA or DNA viruses. cGLR1 is activated by dsRNA to produce the novel CDN 3'2'-cGAMP whereas cGLR2 produces a combination of 2'3'-cGAMP and 3'2' cGAMP in response to a yet unidentified stimulus. Our data establish cGAS as the founding member of a family of receptors sensing different types of nucleic acids and triggering immunity through production of CDNs beyond 2'3'-cGAMP.
Slavik Kailey M, Morehouse Benjamin R, Ragucci Adelyn E, Zhou Wen, Ai Xianlong, Chen Yuqiang, Li Lihua, Wei Ziming, Bähre Heike, König Martin, Seifert Roland, Lee Amy S Y, Cai Hua, Imler Jean-Luc, Kranzusch Philip J
cGAS-like receptors sense RNA and control 3'2'-cGAMP signalling in Drosophila Journal Article
In: Nature, vol. 597, no. 7874, pp. 109–113, 2021, ISSN: 1476-4687.
Abstract | Links | BibTeX | Tags: imler, M3i
@article{pmid34261127,
title = {cGAS-like receptors sense RNA and control 3'2'-cGAMP signalling in Drosophila},
author = {Kailey M Slavik and Benjamin R Morehouse and Adelyn E Ragucci and Wen Zhou and Xianlong Ai and Yuqiang Chen and Lihua Li and Ziming Wei and Heike Bähre and Martin König and Roland Seifert and Amy S Y Lee and Hua Cai and Jean-Luc Imler and Philip J Kranzusch},
doi = {10.1038/s41586-021-03743-5},
issn = {1476-4687},
year = {2021},
date = {2021-07-14},
journal = {Nature},
volume = {597},
number = {7874},
pages = {109--113},
abstract = {Cyclic GMP-AMP synthase (cGAS) is a cytosolic DNA sensor that produces the second messenger cG[2'-5']pA[3'-5']p (2'3'-cGAMP) and controls activation of innate immunity in mammalian cells. Animal genomes typically encode multiple proteins with predicted homology to cGAS, but the function of these uncharacterized enzymes is unknown. Here we show that cGAS-like receptors (cGLRs) are innate immune sensors that are capable of recognizing divergent molecular patterns and catalysing synthesis of distinct nucleotide second messenger signals. Crystal structures of human and insect cGLRs reveal a nucleotidyltransferase signalling core shared with cGAS and a diversified primary ligand-binding surface modified with notable insertions and deletions. We demonstrate that surface remodelling of cGLRs enables altered ligand specificity and used a forward biochemical screen to identify cGLR1 as a double-stranded RNA sensor in the model organism Drosophila melanogaster. We show that RNA recognition activates Drosophila cGLR1 to synthesize the novel product cG[3'-5']pA[2'-5']p (3'2'-cGAMP). A crystal structure of Drosophila stimulator of interferon genes (dSTING) in complex with 3'2'-cGAMP explains selective isomer recognition, and 3'2'-cGAMP induces an enhanced antiviral state in vivo that protects from viral infection. Similar to radiation of Toll-like receptors in pathogen immunity, our results establish cGLRs as a diverse family of metazoan pattern recognition receptors.},
keywords = {imler, M3i},
pubstate = {published},
tppubtype = {article}
}
Cyclic GMP-AMP synthase (cGAS) is a cytosolic DNA sensor that produces the second messenger cG[2'-5']pA[3'-5']p (2'3'-cGAMP) and controls activation of innate immunity in mammalian cells. Animal genomes typically encode multiple proteins with predicted homology to cGAS, but the function of these uncharacterized enzymes is unknown. Here we show that cGAS-like receptors (cGLRs) are innate immune sensors that are capable of recognizing divergent molecular patterns and catalysing synthesis of distinct nucleotide second messenger signals. Crystal structures of human and insect cGLRs reveal a nucleotidyltransferase signalling core shared with cGAS and a diversified primary ligand-binding surface modified with notable insertions and deletions. We demonstrate that surface remodelling of cGLRs enables altered ligand specificity and used a forward biochemical screen to identify cGLR1 as a double-stranded RNA sensor in the model organism Drosophila melanogaster. We show that RNA recognition activates Drosophila cGLR1 to synthesize the novel product cG[3'-5']pA[2'-5']p (3'2'-cGAMP). A crystal structure of Drosophila stimulator of interferon genes (dSTING) in complex with 3'2'-cGAMP explains selective isomer recognition, and 3'2'-cGAMP induces an enhanced antiviral state in vivo that protects from viral infection. Similar to radiation of Toll-like receptors in pathogen immunity, our results establish cGLRs as a diverse family of metazoan pattern recognition receptors.
Cai Hua, Imler Jean-Luc
cGAS-STING: insight on the evolution of a primordial antiviral signaling cassette Journal Article
In: Faculty Reviews, vol. 10, pp. 54, 2021.
Abstract | Links | BibTeX | Tags: imler, M3i
@article{Cai2021,
title = {cGAS-STING: insight on the evolution of a primordial antiviral signaling cassette},
author = {Hua Cai and Jean-Luc Imler
},
url = { https://doi.org/10.12703/r/10-54},
doi = {10.12703/r/10-54},
year = {2021},
date = {2021-06-08},
journal = {Faculty Reviews},
volume = {10},
pages = {54},
abstract = {Stimulator of interferon genes (STING) functions in the cytosolic DNA-sensing pathway of innate immunity in mammals. It is activated upon binding the cyclic dinucleotide 2′3′-cGAMP, a second messenger produced by the enzyme cyclic guanosine monophosphate–adenosine monophosphate synthase (cGAS), which acts as the receptor for DNA in this pathway, and triggers the expression of interferons and other viral stress-induced genes. The ancient origin of STING in the evolution of animals had been noted, but its primitive function was speculative. We review here recent advances in the remarkable history of cGAS-STING signaling, which establish that cGAS is a member of the family of cGAS/DncV-like nucleotidyltransferases (CD-NTases). In bacteria, CD-NTases synthesize a wide range of cyclic oligonucleotide second messengers in response to bacteriophage infections, which in turn activate a variety of effector proteins to abort phage infection. Among these effectors, some are related to STING, revealing an ancestral function for the cGAS-STING cassette in antiviral host defense. Study of STING signaling in invertebrate animals is consistent with an early acquisition in the history of metazoans of CD-NTase- and STING-encoding genes to counter the universal threat of viruses. In particular, STING-dependent immunity appears to play a previously unsuspected important role in some insects. These discoveries open up interesting perspectives for the use of model organisms to decipher emerging aspects of cGAS-STING biology in mammals, such as the activation of interferon-independent responses or the function and regulation of cGAS in the nucleus.},
keywords = {imler, M3i},
pubstate = {published},
tppubtype = {article}
}
Stimulator of interferon genes (STING) functions in the cytosolic DNA-sensing pathway of innate immunity in mammals. It is activated upon binding the cyclic dinucleotide 2′3′-cGAMP, a second messenger produced by the enzyme cyclic guanosine monophosphate–adenosine monophosphate synthase (cGAS), which acts as the receptor for DNA in this pathway, and triggers the expression of interferons and other viral stress-induced genes. The ancient origin of STING in the evolution of animals had been noted, but its primitive function was speculative. We review here recent advances in the remarkable history of cGAS-STING signaling, which establish that cGAS is a member of the family of cGAS/DncV-like nucleotidyltransferases (CD-NTases). In bacteria, CD-NTases synthesize a wide range of cyclic oligonucleotide second messengers in response to bacteriophage infections, which in turn activate a variety of effector proteins to abort phage infection. Among these effectors, some are related to STING, revealing an ancestral function for the cGAS-STING cassette in antiviral host defense. Study of STING signaling in invertebrate animals is consistent with an early acquisition in the history of metazoans of CD-NTase- and STING-encoding genes to counter the universal threat of viruses. In particular, STING-dependent immunity appears to play a previously unsuspected important role in some insects. These discoveries open up interesting perspectives for the use of model organisms to decipher emerging aspects of cGAS-STING biology in mammals, such as the activation of interferon-independent responses or the function and regulation of cGAS in the nucleus.
Leite Thiago H J F, Ferreira Alvaro G A, Imler Jean-Luc, Marques João T
Distinct Roles of Hemocytes at Different Stages of Infection by Dengue and Zika Viruses in Aedes aegypti Mosquitoes Journal Article
In: Frontiers in immunology, vol. 12, pp. 660873, 2021.
Abstract | Links | BibTeX | Tags: Aedes, Dengue, Hemocytes, imler, innate immunity, M3i, Marques, Zika
@article{Leite2021,
title = {Distinct Roles of Hemocytes at Different Stages of Infection by Dengue and Zika Viruses in Aedes aegypti Mosquitoes},
author = {Thiago H J F Leite and Alvaro G A Ferreira and Jean-Luc Imler and João T Marques},
url = {https://www.frontiersin.org/articles/10.3389/fimmu.2021.660873/full},
doi = {10.3389/fimmu.2021.660873},
year = {2021},
date = {2021-05-13},
journal = {Frontiers in immunology},
volume = {12},
pages = {660873},
abstract = {Aedes aegypti mosquitoes are vectors for arboviruses of medical importance such as dengue (DENV) and Zika (ZIKV) viruses. Different innate immune pathways contribute to the control of arboviruses in the mosquito vector including RNA interference, Toll and Jak- STAT pathways. However, the role of cellular responses mediated by circulating macrophage-like cells known as hemocytes remains unclear. Here we show that hemocytes are recruited to the midgut of Ae. aegypti mosquitoes in response to DENV or ZIKV. Blockade of the phagocytic function of hemocytes using latex beads induced increased accumulation of hemocytes in the midgut and a reduction in virus infection levels in this organ. In contrast, inhibition of phagocytosis by hemocytes led to increased systemic dissemination and replication of DENV and ZIKV. Hence, our work reveals a dual role for hemocytes in Ae. aegypti mosquitoes, whereby phagocytosis is not required to control viral infection in the midgut but is essential to restrict systemic dissemination. Further understanding of the mechanism behind this duality could help the design of vector-based strategies to prevent transmission of arboviruses.},
keywords = {Aedes, Dengue, Hemocytes, imler, innate immunity, M3i, Marques, Zika},
pubstate = {published},
tppubtype = {article}
}
Aedes aegypti mosquitoes are vectors for arboviruses of medical importance such as dengue (DENV) and Zika (ZIKV) viruses. Different innate immune pathways contribute to the control of arboviruses in the mosquito vector including RNA interference, Toll and Jak- STAT pathways. However, the role of cellular responses mediated by circulating macrophage-like cells known as hemocytes remains unclear. Here we show that hemocytes are recruited to the midgut of Ae. aegypti mosquitoes in response to DENV or ZIKV. Blockade of the phagocytic function of hemocytes using latex beads induced increased accumulation of hemocytes in the midgut and a reduction in virus infection levels in this organ. In contrast, inhibition of phagocytosis by hemocytes led to increased systemic dissemination and replication of DENV and ZIKV. Hence, our work reveals a dual role for hemocytes in Ae. aegypti mosquitoes, whereby phagocytosis is not required to control viral infection in the midgut but is essential to restrict systemic dissemination. Further understanding of the mechanism behind this duality could help the design of vector-based strategies to prevent transmission of arboviruses.
Chen Di, Roychowdhury-Sinha Arghyashree, Prakash Pragya, Lan Xiao, Fan Wenmin, Goto Akira, Hoffmann Jules A
A time course transcriptomic analysis of host and injected oncogenic cells reveals new aspects of Drosophila immune defenses Journal Article
In: PNAS, vol. 118, no. 12, 2021.
Abstract | Links | BibTeX | Tags: cancer, chemoreceptor, Drosophila melanogaster,