Takeuchi Hiroaki, Saito Hideki, Noda Takeshi, Miyamoto Tadashi, Yoshinaga Tomokazu, Terahara Kazutaka, Ishii Hiroshi, Tsunetsugu-Yokota Yasuko, Yamaoka Shoji
Department of Molecular Virology, Tokyo Medical and Dental University, Tokyo, Japan.
Division of Ultrastructural Virology, International Research Center for Infectious Diseases, Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
PLoS Pathog. 2017 Jul 6;13(7):e1006441. doi: 10.1371/journal.ppat.1006441. eCollection 2017 Jul.
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.
衣壳解聚的调控对于HIV-1进入后高效的cDNA合成至关重要，但参与这一过程的宿主因子仍大多未知。在此，我们利用人类T细胞的遗传筛选来鉴定母源胚胎亮氨酸拉链激酶（MELK），它是HIV-1核心最佳去衣壳化以促进病毒cDNA合成所需的宿主因子。MELK的缺失抑制了HIV-1 cDNA合成，并伴随衣壳解聚延迟。MELK使多聚化HIV-1核心中的衣壳Ser-149位点磷酸化，携带Ser-149磷酸化模拟氨基酸替代的突变病毒发生过早衣壳解聚和更早的HIV-1 cDNA合成，最终无法进入细胞核。此外，一种小分子MELK抑制剂以剂量依赖方式降低了HIV-1在外周血单核细胞中的复制效率。这些结果揭示了一种此前未被认识的HIV-1衣壳解聚机制，并表明MELK是抗HIV治疗的潜在靶点。
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