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2024
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 Article de journal
Dans: BMC Biol , vol. 22, no. 14, 2024.
Résumé | Liens | BibTeX | Étiquettes: 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 Article de journal
Dans: Genetics and Genomics, 2024.
Résumé | Liens | BibTeX | Étiquettes: 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.
2023
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 Article de journal
Dans: Virus Evol., vol. 9, iss. 2, 2023.
Résumé | Liens | BibTeX | Étiquettes: 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.
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 Article de journal
Dans: PLoS Negl. Trop. Dis., vol. 17, 2023.
Résumé | Liens | BibTeX | Étiquettes: 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 Article de journal
Dans: STAR Protocols, vol. 4, iss. 1, 2023.
Résumé | Liens | BibTeX | Étiquettes: 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
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 Article de journal
Dans: Nature Microbiology, vol. 8, iss. 1, 2023.
Résumé | Liens | BibTeX | Étiquettes: 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
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 Article de journal
Dans: Cell Reports, vol. 39, p. 110976, 2022.
Résumé | Liens | BibTeX | Étiquettes: 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.
2021
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 Article de journal
Dans: Frontiers in immunology, vol. 12, p. 660873, 2021.
Résumé | Liens | BibTeX | Étiquettes: 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.
2020
Cai H, Holleufer A, Simonsen B, Schneider J, Lemoine A, Gad HH, Huang J, Huang J, Chen D, Peng T, Marques JT, Hartmann R, Martins N, Imler JL
2'3'-cGAMP triggers a STING- and NF-κB-dependent broad antiviral response in Drosophila Article de journal
Dans: Science Signaling, vol. 13, no. 660, p. eabc4537, 2020.
Résumé | Liens | BibTeX | Étiquettes: imler, M3i, Marques, STING
@article{Cai_2020,
title = {2'3'-cGAMP triggers a STING- and NF-κB-dependent broad antiviral response in Drosophila},
author = {H Cai and A Holleufer and B Simonsen and J Schneider and A Lemoine and HH Gad and J Huang and J Huang and D Chen and T Peng and JT Marques and R Hartmann and N Martins and JL Imler},
url = {https://pubmed.ncbi.nlm.nih.gov/33262294/},
doi = {10.1126/scisignal.abc4537 },
year = {2020},
date = {2020-12-01},
journal = {Science Signaling},
volume = {13},
number = {660},
pages = {eabc4537},
abstract = {We previously reported that an ortholog of STING regulates infection by picorna-like viruses in Drosophila In mammals, STING is activated by the cyclic dinucleotide 2'3'-cGAMP produced by cGAS, which acts as a receptor for cytosolic DNA. Here, we showed that injection of flies with 2'3'-cGAMP induced the expression of dSTING-regulated genes. Coinjection of 2'3'-cGAMP with a panel of RNA or DNA viruses resulted in substantially reduced viral replication. This 2'3'-cGAMP-mediated protection was still observed in flies with mutations in Atg7 and AGO2, genes that encode key components of the autophagy and small interfering RNA pathways, respectively. By contrast, this protection was abrogated in flies with mutations in the gene encoding the NF-κB transcription factor Relish. Transcriptomic analysis of 2'3'-cGAMP-injected flies revealed a complex response pattern in which genes were rapidly induced, induced after a delay, or induced in a sustained manner. Our results reveal that dSTING regulates an NF-κB-dependent antiviral program that predates the emergence of interferons in vertebrates. },
keywords = {imler, M3i, Marques, STING},
pubstate = {published},
tppubtype = {article}
}
We previously reported that an ortholog of STING regulates infection by picorna-like viruses in Drosophila In mammals, STING is activated by the cyclic dinucleotide 2'3'-cGAMP produced by cGAS, which acts as a receptor for cytosolic DNA. Here, we showed that injection of flies with 2'3'-cGAMP induced the expression of dSTING-regulated genes. Coinjection of 2'3'-cGAMP with a panel of RNA or DNA viruses resulted in substantially reduced viral replication. This 2'3'-cGAMP-mediated protection was still observed in flies with mutations in Atg7 and AGO2, genes that encode key components of the autophagy and small interfering RNA pathways, respectively. By contrast, this protection was abrogated in flies with mutations in the gene encoding the NF-κB transcription factor Relish. Transcriptomic analysis of 2'3'-cGAMP-injected flies revealed a complex response pattern in which genes were rapidly induced, induced after a delay, or induced in a sustained manner. Our results reveal that dSTING regulates an NF-κB-dependent antiviral program that predates the emergence of interferons in vertebrates.
Aguiar ERGR, de Almeida JPP, Queiroz LR, Oliveira LS, Olmo RP, de Faria IJDS, Imler JL, Gruber A, Matthews BJ, Marques JT
A single unidirectional piRNA cluster similar to the flamenco locus is the major source of EVE-derived transcription and small RNAs in Aedes aegypti mosquitoes Article de journal
Dans: RNA, vol. 26, no. 5, p. 581-594, 2020.
Résumé | Liens | BibTeX | Étiquettes: A. aegypti, Aedes aegypti, endogenous viral elements, EVE, flamenco locus, imler, M3i, Marques, piRNA, piRNAs, RNA Interference
@article{Aguiar_2020,
title = {A single unidirectional piRNA cluster similar to the flamenco locus is the major source of EVE-derived transcription and small RNAs in Aedes aegypti mosquitoes },
author = {ERGR Aguiar and JPP de Almeida and LR Queiroz and LS Oliveira and RP Olmo and IJDS de Faria and JL Imler and A Gruber and BJ Matthews and JT Marques},
url = {https://rnajournal.cshlp.org/content/26/5/581.long},
doi = {10.1261/rna.073965.119},
year = {2020},
date = {2020-01-29},
journal = {RNA},
volume = {26},
number = {5},
pages = {581-594},
abstract = {Endogenous viral elements (EVEs) are found in many eukaryotic genomes. Despite considerable knowledge about genomic elements such as transposons (TEs) and retroviruses, we still lack information about nonretroviral EVEs. Aedes aegypti mosquitoes have a highly repetitive genome that is covered with EVEs. Here, we identified 129 nonretroviral EVEs in the AaegL5 version of the A. aegypti genome. These EVEs were significantly associated with TEs and preferentially located in repeat-rich clusters within intergenic regions. Genome-wide transcriptome analysis showed that most EVEs generated transcripts although only around 1.4% were sense RNAs. The majority of EVE transcription was antisense and correlated with the generation of EVE-derived small RNAs. A single genomic cluster of EVEs located in a 143 kb repetitive region in chromosome 2 contributed with 42% of antisense transcription and 45% of small RNAs derived from viral elements. This region was enriched for TE-EVE hybrids organized in the same coding strand. These generated a single long antisense transcript that correlated with the generation of phased primary PIWI-interacting RNAs (piRNAs). The putative promoter of this region had a conserved binding site for the transcription factor Cubitus interruptus, a key regulator of the flamenco locus in Drosophila melanogaster Here, we have identified a single unidirectional piRNA cluster in the A. aegypti genome that is the major source of EVE transcription fueling the generation of antisense small RNAs in mosquitoes. We propose that this region is a flamenco-like locus in A. aegypti due to its relatedness to the major unidirectional piRNA cluster in Drosophila melanogaster. },
keywords = {A. aegypti, Aedes aegypti, endogenous viral elements, EVE, flamenco locus, imler, M3i, Marques, piRNA, piRNAs, RNA Interference},
pubstate = {published},
tppubtype = {article}
}
Endogenous viral elements (EVEs) are found in many eukaryotic genomes. Despite considerable knowledge about genomic elements such as transposons (TEs) and retroviruses, we still lack information about nonretroviral EVEs. Aedes aegypti mosquitoes have a highly repetitive genome that is covered with EVEs. Here, we identified 129 nonretroviral EVEs in the AaegL5 version of the A. aegypti genome. These EVEs were significantly associated with TEs and preferentially located in repeat-rich clusters within intergenic regions. Genome-wide transcriptome analysis showed that most EVEs generated transcripts although only around 1.4% were sense RNAs. The majority of EVE transcription was antisense and correlated with the generation of EVE-derived small RNAs. A single genomic cluster of EVEs located in a 143 kb repetitive region in chromosome 2 contributed with 42% of antisense transcription and 45% of small RNAs derived from viral elements. This region was enriched for TE-EVE hybrids organized in the same coding strand. These generated a single long antisense transcript that correlated with the generation of phased primary PIWI-interacting RNAs (piRNAs). The putative promoter of this region had a conserved binding site for the transcription factor Cubitus interruptus, a key regulator of the flamenco locus in Drosophila melanogaster Here, we have identified a single unidirectional piRNA cluster in the A. aegypti genome that is the major source of EVE transcription fueling the generation of antisense small RNAs in mosquitoes. We propose that this region is a flamenco-like locus in A. aegypti due to its relatedness to the major unidirectional piRNA cluster in Drosophila melanogaster.
2019
Olmo RP, Martins NE, Aguiar ERGR, Marques JT, Imler JL
The insect reservoir of biodiversity for viruses and for antiviral mechanisms Article de journal
Dans: An Acad Bras Cienc , vol. 91, no. Suppl 3, p. e20190122, 2019.
Résumé | Liens | BibTeX | Étiquettes: antiviral immunity, imler, insects, M3i, Marques, metagenomics, restriction factors, RNA Interference, virome
@article{Olmo_2019,
title = {The insect reservoir of biodiversity for viruses and for antiviral mechanisms},
author = {RP Olmo and NE Martins and ERGR Aguiar and JT Marques and JL Imler},
url = {https://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652019000600604&lng=en&nrm=iso&tlng=en},
doi = {10.1590/0001-3765201920190122},
year = {2019},
date = {2019-06-03},
journal = { An Acad Bras Cienc },
volume = {91},
number = {Suppl 3},
pages = {e20190122},
abstract = {Insects are the most diverse group of animals. They can be infected by an extraordinary diversity of viruses. Among them, arthropod-borne viruses (arboviruses) can be transmitted to humans. High-throughput sequencing of small RNAs from insects provides insight into their virome, which may help understand the dynamics of vector borne infectious diseases. Furthermore, investigating the mechanisms that restrict viral infections in insects points to genetic innovations that may inspire novel antiviral strategies. },
keywords = {antiviral immunity, imler, insects, M3i, Marques, metagenomics, restriction factors, RNA Interference, virome},
pubstate = {published},
tppubtype = {article}
}
Insects are the most diverse group of animals. They can be infected by an extraordinary diversity of viruses. Among them, arthropod-borne viruses (arboviruses) can be transmitted to humans. High-throughput sequencing of small RNAs from insects provides insight into their virome, which may help understand the dynamics of vector borne infectious diseases. Furthermore, investigating the mechanisms that restrict viral infections in insects points to genetic innovations that may inspire novel antiviral strategies.
Martins NE, Olmo RP, Aguiar ERGR, Marques JT, Imler JL
Les insectes : un fantastique réservoir de virus et de gènes antiviraux Article de journal
Dans: Biologie Aujourd'hui, vol. 212, no. 3-4, p. 101-106, 2019.
Résumé | Liens | BibTeX | Étiquettes: antiviral immunity, ARN interference, imler, Insect, M3i, Marques, metagenomic, virome
@article{Martins_2019jbio,
title = {Les insectes : un fantastique réservoir de virus et de gènes antiviraux},
author = {NE Martins and RP Olmo and ERGR Aguiar and JT Marques and JL Imler},
url = {https://www.biologie-journal.org/articles/jbio/abs/2018/02/jbio190008/jbio190008.html},
doi = {10.1051/jbio/2019008},
year = {2019},
date = {2019-04-11},
journal = {Biologie Aujourd'hui},
volume = {212},
number = {3-4},
pages = {101-106},
abstract = {Les insectes forment le groupe d’animaux qui présente la plus grande diversité. Des travaux récents de métagénomique montrent qu’ils peuvent être infectés par une diversité extraordinaire de virus. Parmi eux, les arbovirus (arthropod-borne viruses) peuvent être transmis à l’Homme par les insectes hématophages, notamment les moustiques. Le séquençage à haut débit des petits ARN des insectes fournit des informations sur leur virome, un paramètre qui pourrait contribuer à expliquer la dynamique de la transmission des maladies infectieuses par des insectes vecteurs. D’autre part, la caractérisation des mécanismes qui restreignent les infections virales chez les insectes révèle des innovations génétiques qui pourraient à terme inspirer de nouvelles stratégies antivirales.},
keywords = {antiviral immunity, ARN interference, imler, Insect, M3i, Marques, metagenomic, virome},
pubstate = {published},
tppubtype = {article}
}
Les insectes forment le groupe d’animaux qui présente la plus grande diversité. Des travaux récents de métagénomique montrent qu’ils peuvent être infectés par une diversité extraordinaire de virus. Parmi eux, les arbovirus (arthropod-borne viruses) peuvent être transmis à l’Homme par les insectes hématophages, notamment les moustiques. Le séquençage à haut débit des petits ARN des insectes fournit des informations sur leur virome, un paramètre qui pourrait contribuer à expliquer la dynamique de la transmission des maladies infectieuses par des insectes vecteurs. D’autre part, la caractérisation des mécanismes qui restreignent les infections virales chez les insectes révèle des innovations génétiques qui pourraient à terme inspirer de nouvelles stratégies antivirales.
2018
Olmo RP, Ferreira AGA, Izidoro-Toledo TC, Aguiar ERGR, de Faria IJS, de Souza KPR, Osório KP, Kuhn L, Hammann P, de Andrade EG, Todjro YM, Rocha MN, Leite THJF, Amadou SCG, Armache JN, Paro S, de Oliveira CD, Carvalho FD, Moreira LA, Marois E, Imler JL, Marques JT
Control of dengue virus in the midgut of Aedes aegypti by ectopic expression of the dsRNA-binding protein Loqs2 Article de journal
Dans: Nature Microbiology, vol. 3, no. 12, p. 1385-1393, 2018.
Résumé | Liens | BibTeX | Étiquettes: Aedes aegypti, Dengue, imler, M3i, marois, Marques, Zika
@article{Olmo_2018,
title = {Control of dengue virus in the midgut of Aedes aegypti by ectopic expression of the dsRNA-binding protein Loqs2 },
author = {RP Olmo and AGA Ferreira and TC Izidoro-Toledo and ERGR Aguiar and IJS de Faria and KPR de Souza and KP Osório and L Kuhn and P Hammann and EG de Andrade and YM Todjro and MN Rocha and THJF Leite and SCG Amadou and JN Armache and S Paro and CD de Oliveira and FD Carvalho and LA Moreira and E Marois and JL Imler and JT Marques},
url = {https://www.nature.com/articles/s41564-018-0268-6},
doi = {10.1038/s41564-018-0268-6},
year = {2018},
date = {2018-10-29},
journal = {Nature Microbiology},
volume = {3},
number = {12},
pages = {1385-1393},
abstract = {Dengue virus (DENV) is an arbovirus transmitted to humans by Aedes mosquitoes. In the insect vector, the small interfering RNA (siRNA) pathway is an important antiviral mechanism against DENV. However, it remains unclear when and where the siRNA pathway acts during the virus cycle. Here, we show that the siRNA pathway fails to efficiently silence DENV in the midgut of Aedes aegypti although it is essential to restrict systemic replication. Accumulation of DENV-derived siRNAs in the midgut reveals that impaired silencing results from a defect downstream of small RNA biogenesis. Notably, silencing triggered by endogenous and exogenous dsRNAs remained effective in the midgut where known components of the siRNA pathway, including the double-stranded RNA (dsRNA)-binding proteins Loquacious and r2d2, had normal expression levels. We identified an Aedes-specific paralogue of loquacious and r2d2, hereafter named loqs2, which is not expressed in the midgut. Loqs2 interacts with Loquacious and r2d2 and is required to control systemic replication of DENV and also Zika virus. Furthermore, ectopic expression of Loqs2 in the midgut of transgenic mosquitoes is sufficient to restrict DENV replication and dissemination. Together, our data reveal a mechanism of tissue-specific regulation of the mosquito siRNA pathway controlled by Loqs2. },
keywords = {Aedes aegypti, Dengue, imler, M3i, marois, Marques, Zika},
pubstate = {published},
tppubtype = {article}
}
Dengue virus (DENV) is an arbovirus transmitted to humans by Aedes mosquitoes. In the insect vector, the small interfering RNA (siRNA) pathway is an important antiviral mechanism against DENV. However, it remains unclear when and where the siRNA pathway acts during the virus cycle. Here, we show that the siRNA pathway fails to efficiently silence DENV in the midgut of Aedes aegypti although it is essential to restrict systemic replication. Accumulation of DENV-derived siRNAs in the midgut reveals that impaired silencing results from a defect downstream of small RNA biogenesis. Notably, silencing triggered by endogenous and exogenous dsRNAs remained effective in the midgut where known components of the siRNA pathway, including the double-stranded RNA (dsRNA)-binding proteins Loquacious and r2d2, had normal expression levels. We identified an Aedes-specific paralogue of loquacious and r2d2, hereafter named loqs2, which is not expressed in the midgut. Loqs2 interacts with Loquacious and r2d2 and is required to control systemic replication of DENV and also Zika virus. Furthermore, ectopic expression of Loqs2 in the midgut of transgenic mosquitoes is sufficient to restrict DENV replication and dissemination. Together, our data reveal a mechanism of tissue-specific regulation of the mosquito siRNA pathway controlled by Loqs2.
Responsable
Tél. :
03 88 41 70 37
Email :
joao.marques@unistra.fr