TY - JOUR T1 - Viral-mediated ubiquitination impacts interactions of host proteins with viral RNA and promotes viral RNA processing JF - bioRxiv DO - 10.1101/2020.06.05.136671 SP - 2020.06.05.136671 AU - Christin Herrmann AU - Joseph M. Dybas AU - Jennifer C. Liddle AU - Alexander M Price AU - Katharina E. Hayer AU - Richard Lauman AU - Caitlin E. Purman AU - Matthew Charman AU - Eui Tae Kim AU - Benjamin A Garcia AU - Matthew D Weitzman Y1 - 2020/01/01 UR - http://biorxiv.org/content/early/2020/06/05/2020.06.05.136671.abstract N2 - Viruses promote infection by hijacking host ubiquitin machinery to counteract or redirect cellular processes. Adenovirus encodes two early proteins, E1B55K and E4orf6, that together co-opt a cellular ubiquitin ligase complex to overcome host defenses and promote virus production. Adenovirus mutants lacking E1B55K or E4orf6 display defects in viral RNA processing and protein production, but previously identified substrates of the redirected ligase do not explain these phenotypes. Here we used a quantitative proteomics approach to identify substrates of E1B55K/E4orf6-mediated ubiquitination that facilitate RNA processing. While all currently known cellular substrates of E1B55K/E4orf6 are degraded by the proteasome, we uncovered RNA-binding proteins (RBPs) as high-confidence substrates which are not decreased in overall abundance. We focused on two RBPs, RALY and hnRNP-C, which we confirm are ubiquitinated without degradation. Knockdown of RALY and hnRNP-C increased levels of viral RNA splicing, protein abundance, and progeny production during infection with E1B55K-deleted virus. Furthermore, infection with virus deleted for E1B55K resulted in increased interaction of hnRNP-C with viral RNA, and attenuation of viral RNA processing. These data suggest viral-mediated ubiquitination of RALY and hnRNP-C relieves a restriction on viral RNA processing, revealing an unexpected role for non-degradative ubiquitination in manipulation of cellular processes during virus infection.Competing Interest StatementThe authors have declared no competing interest. ER -