Article
- The EMBO Journal (2004) 23, 1063 - 1074
- doi:10.1038/sj.emboj.7600123
Published online: 26 February 2004
Subject Category:
Site-specific regulation of the GEF Cdc24p by the scaffold protein Far1p during yeast mating
Philippe Wiget1,2, Yukiko Shimada2,a, Anne-Christine Butty2, Efrei Bi3 and Matthias Peter1,2
- Swiss Federal Institute of Technology Zurich (ETH), Institute of Biochemistry, Zurich, Switzerland
- Swiss Institute for Experimental Cancer Research (ISREC), Chemin des Boveresses 155, Switzerland
- Department of Cell and Developmental Biology, University of Pennsylvania, School of Medicine, Philadelphia, PA, USA
Correspondence to:
Matthias Peter, Swiss Federal Institute of Technology Zurich (ETH), Institute of Biochemistry, ETH Hoenggerberg HPM G 6.2, CH-8093 Zurich, Switzerland. Tel.: +41 1 633 6586; Fax: +41 1 633 1228; E-mail: matthias.peter@bc.biol.ethz.ch
aPresent address: Department of Biochemistry, University of Geneva, 30, quai Ernest-Ansermet, 1211, Geneva 4, Switzerland
Received 2 July 2003; Accepted 19 January 2004
Abstract
Receptor-mediated cell polarization via heterotrimeric G-proteins induces cytoskeletal rearrangements in a variety of organisms. In yeast, Far1p is required for orienting cell growth towards the mating partner by linking activated G
to the guanine-nucleotide exchange factor (GEF) Cdc24p, which activates the Rho-type GTPase Cdc42p. Here we investigated the role of Far1p in the regulation of Cdc24p in vivo. Using time-lapse microscopy of mating cells and artificial membrane targeting of Far1p, we show that Far1p is necessary and sufficient to recruit Cdc24p to the plasma membrane. Wild-type Far1p contains a PH-like domain, which is required for its membrane localization in vivo. Interestingly, expression of membrane-targeted Far1p causes toxicity, most likely by activating Cdc42p uniformly at the cell cortex. The ability of full-length Far1p to function as an activator of Cdc24p in vivo requires its interaction with Cdc24p and G. Our results imply that G not only targets Far1p to the correct site but may also trigger a conformational change in Far1p that is required for its ability to activate Cdc24p in vivo.
Keywords:
- Cdc24p,
- cell polarity,
- Far1p,
- G,
- yeast mating
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