IAV-m6A · Elucidating the role of m6A RNA methylation on the replication and pathogenesis of influenza A virus.
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2017-06-01 → 2020-05-31
- EU contribution
- €264,668
- Participants
- 2
- Scheme
- MSCA-IF-GF
Lines connect the coordinator with its partners.
Results in brief
Elucidating the role of m6A RNA methylation on the replication and pathogenesis of influenza A virus.
Epitranscriptomics is the study of how small marks, or modifications, on RNA molecules affect how they function. The RNA bases are modified by protein 'writers'. These writer proteins are in both eukaryote and prokaryote cells, which tells us that this process of writing modifications onto RNA has been conserved across billions of years of evolution, hinting at their importance. I am interested in determining exactly what these modifications are doing to the RNA in our cells. One way to study this is by looking at how viruses have evolved to use these modifications. In the 1970s and 1980s research was carried out into 1 specific modification, m6A, on influenza A virus. This m6A modification was found to be quite abundant on the influenza RNA, compared to normal human cellular RNA. However, due to limitations in the technologies at the time no further work was performed to investigate what exactly these m6A modifications do. This project focuses on influenza A virus and aims to locate each m6A on each viral RNA. Once I know the location of each m6A modification I will be able to individually remove these sites from viral RNA and investigate exactly how the characteristics of viral RNA are altered due to this loss. I can also use these unmodified viral RNAs to identify whether any cellular RNA-binding proteins specifically only bind regions of RNA containing modifications. By understanding what these modifications do on viral RNAs I can shed some light on their role on cellular RNAs also. In addition, if I find that these modifications are essential for viruses like influenza A, it may be possible to design antiviral drugs to stop these modifications being added to influenza RNA and thus block infections. The overall objectives of this work are to; locate where each of these m6A modifications are on influenza A RNAs, define exactly how these modifications affect their target RNA and the cell as a whole, work out whether influenza somehow regulates the amount of modifications happening in a cell after infection, and to check whether these m6A modifications result in the specific recruitment of some cellular RNA binding proteins to the site of modification.
Data: CORDIS, © European Union
Project objective
It has recently become apparent that post-transcriptional editing of mRNA by methylation of adenosines (m6A) plays a key role in regulating gene expression. In particular, total loss of m6A editing blocks the development of early embryos in a number of organisms. However, how m6A modifications, which are added in the nucleus and primarily detected by 3 cytoplasmic “reader” proteins, exert their effects remains unknown. The presence of m6A on viral transcripts has been known for 40 years, yet has only recently been shown to enhance HIV-1 gene expression and replication. Influenza A virus (IAV) that, like HIV-1, replicates in the nucleus, is known to bear a high level of m6A on viral RNA. However, whether and how m6A editing affects IAV replication remains unknown. Here, the researcher proposes the investigation of the role of m6A in IAV pathogenesis. The main objectives of this proposal are; to compile a precise map of m6A sites on IAV-PR8 RNA, determine their functional significance on replication and RNA trafficking, assess how m6A machinery is altered in IAV infected cells, and evaluate the conservation of these sites across additional pathogenic IAV strains.Preliminary data demonstrates that IAV gene expression may be enhanced by overexpression of the m6A reader proteins. Therefore the researcher proposes that IAV may subvert the m6A modification pathway to enhance viral RNA expression. Mapping of m6A modifications will be performed using PAR-CLIP and PA-m6A-seq, while RNAi, CRISPR knockouts and lentiviral overexpression systems will be utilised to determine the functional role of m6A editing on IAV pathogenesis. This fellowship will provide the researcher with an excellent opportunity to work at 2 highly regarded research institutes and advance his knowledge in virology and RNA biology. The skills acquired through completion of this fellowship will greatly assist him in his future endeavours including the establishment of his own European research group.
Original text from CORDIS.
Participants
- INSTITUT PASTEUR · ParisCoordinatorFrance
- DUKE UNIVERSITY · Durham NcUnited States
Links
Data: CORDIS, © European Union