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. 2006 Sep 22;78(11):1365–1373. doi: 10.1002/jmv.20707

SARS‐CoV nucleocapsid protein binds to hUbc9, a ubiquitin conjugating enzyme of the sumoylation system

Zheng Fan 1, Yue Zhuo 1,2, Xinyu Tan 3, Zhi Zhou 1,4, Jiangang Yuan 3, Boqin Qiang 3, Jinghua Yan 1, Xiaozhong Peng 3, George F Gao 1,✉
PMCID: PMC7167196  PMID: 16998888

Abstract

SARS‐CoV is a newly identified coronavirus (CoV) that causes severe acute respiratory syndrome (SARS). The SARS‐CoV nucleocapsid (N) protein is an important structural and functional protein. To identify cellular proteins that interact with the SARS‐CoV N protein and to elucidate the possible involvement of N protein in SARS‐CoV pathogenesis, a human lymphocyte cDNA library was screened using a yeast two‐hybrid system assay. hUbc9, a ubiquitin conjugating enzyme of sumoylation system, was found to interact specifically with the N protein, implying the post‐translational sumoylation of the N protein. Mapping studies localized the critical N sequences for this interaction to amino acids 170–210, which includes the SR‐rich motif. However, the consensus motif of sumoylation GK62EE in the N protein is not responsible for binding to hUbc9. Mutations of hUbc9 at the enzyme active site C93A or C93S severely impair the interaction with the N protein. The two proteins were also shown to colocalize in the cytoplasm of the transfected 293T cells. This is the first report demonstrating the interaction of hUbc9 with a structural protein of plus‐strand RNA viruses, indicating a new drug target for SARS‐CoV. J. Med. Virol. 78:1365–1373, 2006. © 2006 Wiley‐Liss, Inc.

Keywords: severe acute respiratory syndrome, coronavirus, SARS‐CoV, nucleocapsid (N) protein, hUbc9, interaction

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