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

• Published in: PLoS pathogens 2017-07, Vol.13 (7), p.e1006441-e1006441
• Main Authors: Takeuchi, Hiroaki, Saito, Hideki, Noda, Takeshi, Miyamoto, Tadashi, Yoshinaga, Tomokazu, Terahara, Kazutaka, Ishii, Hiroshi, Tsunetsugu-Yokota, Yasuko, Yamaoka, Shoji
• Format: Article
• Language: English
• Subjects: Acquired immune deficiency syndrome; AIDS; Amino acid substitution; Amino acids; Biology and Life Sciences; Capsid - metabolism; Cell Line; Chemical synthesis; Dismantling; DNA, Viral - genetics; DNA, Viral - metabolism; Efficiency; Genetic aspects; Genetic screening; Health aspects; HIV; HIV Infections - enzymology; HIV Infections - genetics; HIV Infections - virology; HIV-1 - genetics; HIV-1 - physiology; Host-Pathogen Interactions; Human immunodeficiency virus; Humans; Immunology; Infections; Infectious diseases; Laboratories; Leucine; Leucine zipper proteins; Leukemia; Leukocytes (mononuclear); Leukocytes, Mononuclear - enzymology; Leukocytes, Mononuclear - virology; Lymphocytes T; Medical research; Medicine and Health Sciences; Mutation; Peripheral blood mononuclear cells; Phosphorylation; Phosphotransferases; Physical Sciences; Protein-Serine-Threonine Kinases - genetics; Protein-Serine-Threonine Kinases - metabolism; Proteins; R&D; Research & development; Research and analysis methods; Science; Uncoating; Virology; Virulence (Microbiology); Virus Replication; Virus Uncoating; Viruses
• ISSN: 1553-7374; 1553-7366; 1553-7374
• DOI: 10.1371/journal.ppat.1006441

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.