The yeast exchange assay, a new complementary method to screen for Db1-like protein specificity: identification of a novel RhoA exchange factor

被引：15
作者：
De Toledo, M
Colombo, K
Nagase, T
Ohara, O
Fort, P
Blangy, A
机构：
[1] Ctr Rech Biochim Macromol, CNRS, UPR 1086, F-34293 Montpellier 5, France
[2] Kazusa DNA Res Inst, Chiba 292, Japan
来源：
FEBS LETTERS | 2000年 / 480卷 / 2-3期
关键词：
Rho GTPase; exchange factor; functional assay;
D O I：
10.1016/S0014-5793(00)01953-0
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：
The target Rho GTPases of many guanine nucleotide exchange factors (GEFs) of the Dbl family remain to be identified. Here we report a new method: the yeast exchange assay (YEA), a rapid qualitative test to perform a wide range screen for GEF specificity. In this assay based on the two-hybrid system, a wild type GTPase binds to its effector only after activation by a specific GEF, We validated the YEA by activating GTPases by previously reported GEFs, We further established that a novel GEF, GEF337, activates RhoA in the YEA. GEF337 promoted nucleotide exchange on RhoA in vitro and promoted F-actin stress fiber assembly in fibroblasts, characteristic of RhoA activation, (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.
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收藏
页码：287 / 292
页数：6
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