Phosphorylation of the HIV-1 capsid by MELK triggers uncoating to promote viral cDNA synthesis

PLoS Pathogens

Shoji Yamaoka¹  Yasuko Tsunetsugu-Yokota²  Hiroaki Takeuchi³  Hideki Saito³  Kazutaka Terahara⁴  Hiroshi Ishii⁴  Takeshi Noda⁵  Tomokazu Yoshinaga⁶  Tadashi Miyamoto⁷ 

[1] AIDS Research Center, National Institute of Infectious Diseases, Tokyo, Japan;Department of Immunology, National Institute of Infectious Diseases, Tokyo, Japan;Department of Molecular Virology, Tokyo Medical and Dental University, Tokyo, Japan;Discovery Research Laboratory for Core Therapeutic Areas, Shionogi Pharmaceutical Research Center, Shionogi & CO., LTD, Osaka, Japan;Division of Ultrastructural Virology, International Research Center for Infectious Diseases, Institute of Medical Science, The University of Tokyo, Tokyo, Japan;Laboratory of Ultrastructural Virology, Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan;PRESTO, Japan Science and Technology Agency, Saitama, Japan

关键词: HIV-1;    Phosphorylation;    cDNA synthesis;    Viral core;    Analysis of variance;    Enzyme-linked immunoassays;    Viral packaging;    In vitro kinase assay;   

DOI  :  10.1371/journal.ppat.1006441

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Regulation of capsid disassembly is crucial for efficient HIV-1 cDNA synthesis after entry, yet host factors involved in this process remain largely unknown. Here, we employ genetic screening of human T-cells to identify maternal embryonic leucine zipper kinase (MELK) as a host factor required for optimal uncoating of the HIV-1 core to promote viral cDNA synthesis. Depletion of MELK inhibited HIV-1 cDNA synthesis with a concomitant delay of capsid disassembly. MELK phosphorylated Ser-149 of the capsid in the multimerized HIV-1 core, and a mutant virus carrying a phosphorylation-mimetic amino-acid substitution of Ser-149 underwent premature capsid disassembly and earlier HIV-1 cDNA synthesis, and eventually failed to enter the nucleus. Moreover, a small-molecule MELK inhibitor reduced the efficiency of HIV-1 replication in peripheral blood mononuclear cells in a dose-dependent manner. These results reveal a previously unrecognized mechanism of HIV-1 capsid disassembly and implicate MELK as a potential target for anti-HIV therapy.

【 授权许可】

CC BY