Rac1 GTPase activates the WAVE regulatory complex through two distinct binding sites

被引：97

作者：

Chen, Baoyu ^{[1
,2
,5
]}

Chou, Hui-Ting ^{[3
,7
]}

Brautigam, Chad A. ^{[1
,4
]}

Xing, Wenmin ^{[1
,2
]}

Yang, Sheng ^{[5
]}

Henry, Lisa ^{[1
,2
]}

Doolittle, Lynda K. ^{[1
,2
]}

Walz, Thomas ^{[6
]}

Rosen, Michael K. ^{[1
,2
]}

机构：

[1] Univ Texas Southwestern Med Ctr Dallas, Dept Biophys, Dallas, TX 75390 USA

[2] Univ Texas Southwestern Med Ctr Dallas, Howard Hughes Med Inst, Dallas, TX 75390 USA

[3] Harvard Med Sch, Dept Cell Biol, Boston, MA USA

[4] Univ Texas Southwestern Med Ctr Dallas, Dept Microbiol, Dallas, TX USA

[5] Iowa State Univ, Roy J Carver Dept Biochem Biophys & Mol Biol, Ames, IA 50011 USA

[6] Rockefeller Univ, 1230 York Ave, New York, NY 10021 USA

[7] Howard Hughes Med Inst, CryoEM Shared Resources, Jaelia Res Campus, Ashburn, VA USA

来源：

ELIFE | 2017年 / 6卷

基金：

美国国家卫生研究院;

关键词：

VELOCITY ANALYTICAL ULTRACENTRIFUGATION; ALDRICH-SYNDROME PROTEIN; ACTIN NUCLEATION; TYROSINE KINASE; CRYO-EM; ELECTRON-MICROSCOPY; ARP2/3; COMPLEX; WASP; CYTOSKELETON; MEMBRANE;

D O I：

10.7554/eLife.29795

中图分类号：

Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

The Rho GTPase Rac1 activates the WAVE regulatory complex (WRC) to drive Arp2/3 complex-mediated actin polymerization, which underpins diverse cellular processes. Here we report the structure of a WRC-Rac1 complex determined by cryo-electron microscopy. Surprisingly, Rac1 is not located at the binding site on the Sra1 subunit of the WRC previously identified by mutagenesis and biochemical data. Rather, it binds to a distinct, conserved site on the opposite end of Sra1. Biophysical and biochemical data on WRC mutants confirm that Rac1 binds to both sites, with the newly identified site having higher affinity and both sites required for WRC activation. Our data reveal that the WRC is activated by simultaneous engagement of two Rac1 molecules, suggesting a mechanism by which cells may sense the density of active Rac1 at membranes to precisely control actin assembly.

引用

页数：22

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