Poster abstracts
Poster number 102 submitted by Christina Ross
Site-specific identification of m6A RNA modification in HIV-1 genomic RNA 5'UTR
Christina Ross (Molecular, Cellular and Developmental Biology, Department of Chemistry and Biochemistry, Center for Retrovirus Research, and Center for RNA Biology, The Ohio State University, Columbus, Ohio 43210), Karin Musier-Forsyth (Department of Chemistry and Biochemistry, Center for Retrovirus Research, and Center for RNA Biology, The Ohio State University, Columbus, Ohio 43210)
Abstract:
RNA modifications affect all aspects of RNA biology, likely impacting processes in viral replication such as RNA localization, stability, and assembly. To assemble virions, HIV-1 must select full-length genomic RNA (gRNA) from the cytoplasmic pool of cellular and viral RNA. The viral polyprotein Gag orchestrates the packaging process via specific interactions between within the gRNA 5'UTR. Two N-6-methyladenosine (m6A) modifications have been previously identified in the 5'UTR using low resolution m6A-sequencing-based techniques. One of these modifications is located in the viral RNA packaging signal, proximal to a known site of Gag interaction. The presence of both modifications has recently been proposed to play a critical role in down-regulating gRNA packaging. Thus, we hypothesize m6A modifications in the HIV-1 5'UTR influence viral replication and assembly through modulating RNA structure and Gag interactions. Using an approach related to SCARLET (site-specific cleavage and radioactive-labeling followed by ligation-assisted extraction and thin-layer chromatography), we purified RNA from cells transfected with HIV-1 plasmid to determine the stoichiometry of m6A on the specific sites in the gRNA 5'UTR . Future work will investigate the impact of m6A on gRNA packaging by determining the levels of m6A at the 5'UTR sites on cellular versus packaged gRNA.
Keywords: HIV-1, m6A, RNA