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Article Dans Une Revue FEBS Letters Année : 2000

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

Résumé

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.

Dates et versions

hal-02267482 , version 1 (19-08-2019)

Identifiants

Citer

M. de Toledo, K. Colombo, T. Nagase, O. Ohara, Philippe Fort, et al.. The yeast exchange assay, a new complementary method to screen for Dbl-like protein specificity: identification of a novel RhoA exchange factor. FEBS Letters, 2000, 480 (2-3), pp.287-292. ⟨10.1016/s0014-5793(00)01953-0⟩. ⟨hal-02267482⟩
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