Interaction with the Src Homology (SH3-SH2) Region of the Src-family Kinase Hck Structures the HIV-1 Nef Dimer for Kinase Activation and Effector Recruitment

被引:36
作者
Alvarado, John Jeff [1 ,2 ]
Tarafdar, Sreya [1 ,3 ]
Yeh, Joanne I. [2 ]
Smithgall, Thomas E. [1 ]
机构
[1] Univ Pittsburgh, Sch Med, Dept Microbiol & Mol Genet, Pittsburgh, PA 15219 USA
[2] Univ Pittsburgh, Sch Med, Dept Biol Struct, Pittsburgh, PA 15219 USA
[3] Univ Pittsburgh, Grad Sch Publ Hlth, Dept Infect Dis & Microbiol, Pittsburgh, PA 15261 USA
基金
美国国家卫生研究院;
关键词
HUMAN-IMMUNODEFICIENCY-VIRUS; CRYSTAL-STRUCTURE; TYROSINE KINASE; SH3; DOMAIN; DOWN-REGULATION; TYPE-1; NEF; C-SRC; PROTEIN; BINDING; SURFACE;
D O I
10.1074/jbc.M114.600031
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
HIV-1 Nef supports high titer viral replication in vivo and is essential for AIDS progression. Nef function depends on interactions with multiple host cell effectors, including Hck and other Src-family kinases. Here we describe the x-ray crystal structure of Nef in complex with the Hck SH3-SH2 regulatory region to a resolution of 1.86 angstrom. The complex crystallized as a dimer of complexes, with the conserved Nef PXXPXR motif engaging the Hck SH3 domain. Anew intercomplex contact was found between SH3 Glu-93, and Nef Arg-105. Mutagenesis of Hck SH3 Glu-93 interfered with Nef.Hck complex formation and kinase activation in cells. The Hck SH2 domains impinge on the N-terminal region of Nef to stabilize a dimer conformation that exposes Asp-123, a residue critical for Nef function. Our results suggest that in addition to serving as a kinase effector for Nef, Hck binding may reorganize the Nef dimer for functional interaction with other signaling partners.
引用
收藏
页码:28539 / 28553
页数:15
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