基因组
寄主(生物学)
鉴定(生物学)
生物
甲型肝炎病毒
病毒学
计算生物学
乙型肝炎病毒
病毒
宿主因子
遗传学
基因
植物
作者
Isabella T. Whitworth,Sofia Romero,Abena Kissi-Twum,Rachel A. Knoener,Mark Scalf,Nathan M. Sherer,Lloyd M. Smith
标识
DOI:10.1021/acs.jproteome.4c00505
摘要
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.
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