The yeast exchange assay, a new complementary method to screen for Dbl-like protein specificity: identification of a novel RhoA exchange factor
In: FEBS Letters, , Heft 2-3, S. 287-292
Online
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Zugriff:
International audience; 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.
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The yeast exchange assay, a new complementary method to screen for Dbl-like protein specificity: identification of a novel RhoA exchange factor
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| Autor/in / Beteiligte Person: | Fort, Philippe ; Blangy, Anne ; Ohara, Osamu ; M. De Toledo ; Nagase, Takahiro ; Colombo, K. ; Centre de recherche en Biologie Cellulaire (CRBM) ; Université Montpellier 1 (UM1)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Université Montpellier 2 - Sciences et Techniques (UM2) ; Institute of Mineralogy, Petrology, and Economic Geology ; Graduate School of Science ; Centre National de la Recherche Scientifique (CNRS) ; Centre de recherches de biochimie macromoléculaire (CRBM) ; Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-IFR122-Centre National de la Recherche Scientifique (CNRS) ; Centre National de la Recherche Scientifique (CNRS)-IFR122-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Montpellier 1 (UM1) ; Kazusa DNA Research Institute (KDRI) |
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| Zeitschrift: | FEBS Letters, , Heft 2-3, S. 287-292 |
| Veröffentlichung: | Federation of European Biochemical Societies. Published by Elsevier B.V. |
| Medientyp: | unknown |
| ISSN: | 0014-5793 (print) ; 1873-3468 (print) |
| DOI: | 10.1016/S0014-5793(00)01953-0 |
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