| Autor: |
Whitworth IT; Department of Chemistry, University of Wisconsin-Madison College of Letters and Sciences, Madison, Wisconsin 53706, United States., Romero S; McArdle Laboratory for Cancer Research, University of Wisconsin-Madison School of Medicine and Public Health, Madison, Wisconsin 53705, United States.; Institute for Molecular Virology, University of Wisconsin-Madison, Madison, Wisconsin 53706, United States.; Microbiology Doctoral Training Program, University of Wisconsin-Madison, Madison, Wisconsin 53706, United States., Kissi-Twum A; McArdle Laboratory for Cancer Research, University of Wisconsin-Madison School of Medicine and Public Health, Madison, Wisconsin 53705, United States.; Microbiology Doctoral Training Program, University of Wisconsin-Madison, Madison, Wisconsin 53706, United States., Knoener R; Department of Chemistry, University of Wisconsin-Madison College of Letters and Sciences, Madison, Wisconsin 53706, United States.; McArdle Laboratory for Cancer Research, University of Wisconsin-Madison School of Medicine and Public Health, Madison, Wisconsin 53705, United States.; Institute for Molecular Virology, University of Wisconsin-Madison, Madison, Wisconsin 53706, United States., Scalf M; Department of Chemistry, University of Wisconsin-Madison College of Letters and Sciences, Madison, Wisconsin 53706, United States., Sherer NM; McArdle Laboratory for Cancer Research, University of Wisconsin-Madison School of Medicine and Public Health, Madison, Wisconsin 53705, United States.; Institute for Molecular Virology, University of Wisconsin-Madison, Madison, Wisconsin 53706, United States., Smith LM; Department of Chemistry, University of Wisconsin-Madison College of Letters and Sciences, Madison, Wisconsin 53706, United States. |
| Abstrakt: |
A critical part of the hepatitis B virus (HBV) life cycle is the packaging of the pregenomic RNA (pgRNA) into nucleocapsids. While this process is known to involve several viral elements, much less is known about the identities and roles of host proteins in this process. To better understand the role of host proteins, we isolated pgRNA and characterized its protein interactome in cells expressing either packaging-competent or packaging-incompetent HBV genomes. We identified over 250 host proteins preferentially associated with pgRNA from the packaging-competent version of the virus. These included proteins already known to support capsid formation, enhance viral gene expression, catalyze nucleocapsid dephosphorylation, and bind to the viral genome, demonstrating the ability of the approach to effectively reveal functionally significant host-virus interactors. Three of these host proteins, AURKA, YTHDF2, and ATR, were selected for follow-up analysis. RNA immunoprecipitation qPCR (RIP-qPCR) confirmed pgRNA-protein association in cells, and siRNA knockdown of the proteins showed decreased encapsidation efficiency. This study provides a template for the use of comparative RNA-protein interactome analysis in conjunction with virus engineering to reveal functionally significant host-virus interactions. |
