The Cellular Interactome of the Coronavirus Infectious Bronchitis Virus Nucleocapsid Protein and Functional Implications for Virus Biology

• DOI: 10.1128/jvi.00321-13
• Publication Date: 2013-05-02T08:36:26Z
• Authors: Edward Emmott; Diane Munday; Erica Bickerton; Paul Britton; Mark A. Rodgers; Adrian Whitehouse; En-Min Zhou; Julian A. Hiscox
• Keywords: Interactome; Coronavirus; Coronaviridae; Sars virus; 2019-20 coronavirus outbreak

ABSTRACT The coronavirus nucleocapsid (N) protein plays a multifunctional role in the virus life cycle, from regulation of replication and transcription genome packaging to modulation host cell processes. These functions are likely be facilitated by interactions with proteins. potential interactome infectious bronchitis (IBV) N was mapped using stable isotope labeling amino acids culture (SILAC) coupled green fluorescent protein-nanotrap pulldown methodology liquid chromatography-tandem mass spectrometry. addition SILAC label allowed discrimination proteins that were specifically bind over background binding. Overall, 142 cellular selected as potentially binding protein, many part larger possible complexes. included ribosomal proteins, nucleolar translation initiation factors, helicases, hnRNPs. association IBV confirmed immunoblotting, cosedimentation, confocal microscopy. Further, localization IBV-infected cells well their activity during infection assessed small interfering RNA-mediated depletion, demonstrating functional importance biology IBV. This not only confirms previous observations made other both overexpressed but also provides novel data can exploited understand interaction between cell.