Publications
2018
Smyth R. P., Smith M. R., Jousset A. C., Despons L., Laumond G., Decoville T., Cattenoz P., Moog C., Jossinet F., Mougel M., Paillart J. C., Kleist M., Marquet R.
Dans: Nucleic Acids Res, vol. 46, no. 9, p. e57, 2018, ISBN: 29514260, (1362-4962 (Electronic) 0305-1048 (Linking) Journal Article Research Support, Non-U.S. Gov't).
Résumé | Liens | BibTeX | Étiquettes: 5' Untranslated Regions Genome, LESCURE, Ribonucleic Acid *Virus Assembly Virus Replication, Unité ARN, Viral HEK293 Cells HIV-1/*genetics/physiology Humans Mutation Nucleotide Motifs Poly A/metabolism RNA, Viral/*biosynthesis/*chemistry *Regulatory Sequences
@article{nokey,
title = {In cell mutational interference mapping experiment (in cell MIME) identifies the 5' polyadenylation signal as a dual regulator of HIV-1 genomic RNA production and packaging},
author = {R. P. Smyth and M. R. Smith and A. C. Jousset and L. Despons and G. Laumond and T. Decoville and P. Cattenoz and C. Moog and F. Jossinet and M. Mougel and J. C. Paillart and M. Kleist and R. Marquet},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=29514260},
doi = {10.1093/nar/gky152},
isbn = {29514260},
year = {2018},
date = {2018-01-01},
journal = {Nucleic Acids Res},
volume = {46},
number = {9},
pages = {e57},
abstract = {Non-coding RNA regulatory elements are important for viral replication, making them promising targets for therapeutic intervention. However, regulatory RNA is challenging to detect and characterise using classical structure-function assays. Here, we present in cell Mutational Interference Mapping Experiment (in cell MIME) as a way to define RNA regulatory landscapes at single nucleotide resolution under native conditions. In cell MIME is based on (i) random mutation of an RNA target, (ii) expression of mutated RNA in cells, (iii) physical separation of RNA into functional and non-functional populations, and (iv) high-throughput sequencing to identify mutations affecting function. We used in cell MIME to define RNA elements within the 5' region of the HIV-1 genomic RNA (gRNA) that are important for viral replication in cells. We identified three distinct RNA motifs controlling intracellular gRNA production, and two distinct motifs required for gRNA packaging into virions. Our analysis reveals the 73AAUAAA78 polyadenylation motif within the 5' PolyA domain as a dual regulator of gRNA production and gRNA packaging, and demonstrates that a functional polyadenylation signal is required for viral packaging even though it negatively affects gRNA production.},
note = {1362-4962 (Electronic)
0305-1048 (Linking)
Journal Article
Research Support, Non-U.S. Gov't},
keywords = {5' Untranslated Regions Genome, LESCURE, Ribonucleic Acid *Virus Assembly Virus Replication, Unité ARN, Viral HEK293 Cells HIV-1/*genetics/physiology Humans Mutation Nucleotide Motifs Poly A/metabolism RNA, Viral/*biosynthesis/*chemistry *Regulatory Sequences},
pubstate = {published},
tppubtype = {article}
}
Non-coding RNA regulatory elements are important for viral replication, making them promising targets for therapeutic intervention. However, regulatory RNA is challenging to detect and characterise using classical structure-function assays. Here, we present in cell Mutational Interference Mapping Experiment (in cell MIME) as a way to define RNA regulatory landscapes at single nucleotide resolution under native conditions. In cell MIME is based on (i) random mutation of an RNA target, (ii) expression of mutated RNA in cells, (iii) physical separation of RNA into functional and non-functional populations, and (iv) high-throughput sequencing to identify mutations affecting function. We used in cell MIME to define RNA elements within the 5' region of the HIV-1 genomic RNA (gRNA) that are important for viral replication in cells. We identified three distinct RNA motifs controlling intracellular gRNA production, and two distinct motifs required for gRNA packaging into virions. Our analysis reveals the 73AAUAAA78 polyadenylation motif within the 5' PolyA domain as a dual regulator of gRNA production and gRNA packaging, and demonstrates that a functional polyadenylation signal is required for viral packaging even though it negatively affects gRNA production.