The phosphorylation of HIV-1 Gag by atypical protein kinase C facilitates viral infectivity by promoting Vpr incorporation into virions

被引:28
作者
Kudoh, Ayumi [1 ]
Takahama, Shoukichi [2 ]
Sawasaki, Tatsuya [2 ,3 ]
Ode, Hirotaka [4 ,5 ]
Yokoyama, Masaru [5 ]
Okayama, Akiko [6 ]
Ishikawa, Akiyo [6 ]
Miyakawa, Kei [1 ]
Matsunaga, Satoko [1 ]
Kimura, Hirokazu [7 ]
Sugiura, Wataru [4 ]
Sato, Hironori [5 ]
Hirano, Hisashi [6 ]
Ohno, Shigeo [8 ]
Yamamoto, Naoki [9 ]
Ryo, Akihide [1 ]
机构
[1] Yokohama City Univ, Sch Med, Dept Microbiol, Yokohama, Kanagawa 232, Japan
[2] Ehime Univ, Venture Business Lab, Matsuyama, Ehime, Japan
[3] Ehime Univ, Proteosci Ctr, Matsuyama, Ehime, Japan
[4] Natl Hosp Org Nagoya Med Ctr, Clin Res Ctr, Nagoya, Aichi, Japan
[5] Natl Inst Infect Dis, Pathogen Genom Ctr, Tokyo, Japan
[6] Yokohama City Univ, Int Grad Sch Arts & Sci, Yokohama, Kanagawa 232, Japan
[7] Natl Inst Infect Dis, Infect Dis Surveillance Ctr, Tokyo, Japan
[8] Yokohama City Univ, Sch Med, Dept Mol Biol, Yokohama, Kanagawa 232, Japan
[9] Natl Univ Singapore, Dept Microbiol, Singapore 117548, Singapore
来源
RETROVIROLOGY | 2014年 / 11卷
关键词
HIV-1; infection; Phosphorylation; Vpr; aPKC; HUMAN-IMMUNODEFICIENCY-VIRUS; CELL-CYCLE ARREST; MUTATIONAL ANALYSIS; LATE-DOMAIN; TYPE-1; VPR; SYNTHESIS SYSTEM; P6; REPLICATION; RELEASE; TRANSCRIPTION;
D O I
10.1186/1742-4690-11-9
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Background: Human immunodeficiency virus type 1 (HIV-1) Gag is the main structural protein that mediates the assembly and release of virus-like particles (VLPs) from an infected cell membrane. The Gag C-terminal p6 domain contains short sequence motifs that facilitate virus release from the plasma membrane and mediate incorporation of the viral Vpr protein. Gag p6 has also been found to be phosphorylated during HIV-1 infection and this event may affect virus replication. However, the kinase that directs the phosphorylation of Gag p6 toward virus replication remains to be identified. In our present study, we identified this kinase using a proteomic approach and further delineate its role in HIV-1 replication. Results: A proteomic approach was designed to systematically identify human protein kinases that potently interact with HIV-1 Gag and successfully identified 22 candidates. Among this panel, atypical protein kinase C (aPKC) was found to phosphorylate HIV-1 Gag p6. Subsequent LC-MS/MS and immunoblotting analysis with a phospho-specific antibody confirmed both in vitro and in vivo that aPKC phosphorylates HIV-1 Gag at Ser487. Computer-assisted structural modeling and a subsequent cell-based assay revealed that this phosphorylation event is necessary for the interaction between Gag and Vpr and results in the incorporation of Vpr into virions. Moreover, the inhibition of aPKC activity reduced the Vpr levels in virions and impaired HIV-1 infectivity of human primary macrophages. Conclusion: Our current results indicate for the first time that HIV-1 Gag phosphorylation on Ser487 is mediated by aPKC and that this kinase may regulate the incorporation of Vpr into HIV-1 virions and thereby supports virus infectivity. Furthermore, aPKC inhibition efficiently suppresses HIV-1 infectivity in macrophages. aPKC may therefore be an intriguing therapeutic target for HIV-1 infection.
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页数:16
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