Poster abstracts
Poster number 83 submitted by Vidusha Kothwela
Orthogonal Organic Phase Separation (OOPS) Reveals Cellular RNA Binding Proteome Changes in Response to Flavivirus Infections
Vidusha Kothwela (Case Western Reserve University, School of Medicine, Biochemistry Department, Cleveland, OH), Joseph Luna (Case Western Reserve University, School of Medicine, Biochemistry Department, Cleveland, OH)
Abstract:
RNA viruses in the Flaviviridae family, such as Zika virus (ZIKV) and Hepatitis C virus (HCV), heavily rely on host RNA-binding proteins (RBPs) throughout their life cycle. These viruses present a significant global health challenge, with effective antiviral therapies remaining elusive. Our limited understanding of how cellular RBPs influence ZIKV and HCV infections hinders progress toward effective treatments. By investigating RBPome remodeling during these infections, we aim to uncover crucial insights into how RBPs regulate viral replication and immune evasion. We used Orthogonal Organic Phase Separation (OOPS) to profile the RNA-bound proteome during ZIKV and HCV infections. Our analysis revealed virus-specific alterations in RNA-binding activity, with 115 RBPs identified in ZIKV-infected cells and 135 RBPs in HCV-infected cells. These RBPs were distinct from those previously reported for both viruses, suggesting a specialized and dynamic role for RBPs during infection. Only five RBPs exhibited increased RNA-binding activity in both infections, and these did not follow a discernible pattern of functionality, underscoring the specificity of the RBP response to each virus. Among the RBPs identified, the ATP-binding RNA helicase DHX38 showed a marked increase in RNA binding upon ZIKV and HCV infections. Known for its role in pre-mRNA splicing as part of the spliceosome, DHX38 did not exhibit transcriptional activation during infection, suggesting a novel, non-canonical function in viral infection. These findings demonstrate OOPS's robust and broadly applicable nature in uncovering novel RNA-protein interactions during RNA virus infections. More importantly, they provide new insights into how ZIKV and HCV remodel the RBPome, offering potential targets for therapeutic intervention.
Keywords: OOPS, RBPome, Flavivirus