title: Evidence for specific interaction between the RhoGAP domain from the yeast Rgd1 protein and phosphoinositides.
abstract: The Rho GTPase activating protein Rgd1 increases the GTPase activity of Rho3p and Rho4p, which are involved in bud growth and cytokinesis, respectively, in the budding yeast Saccharomyces cerevisiae. Rgd1p is a member of the F-BAR family conserved in eukaryotes; indeed, in addition to the C-terminal RhoGAP domain Rgd1p possesses an F-BAR domain at its N-terminus. Phosphoinositides discriminate between the GTPase activities of Rho3p and Rho4p through Rgd1p and specifically stimulate the RhoGAP activity of Rgd1p on Rho4p. Determining specific interactions and resolving the structure of Rgd1p should provide insight into the functioning of this family of protein. We report the preparation of highly pure and functional RhoGAP domain of Rgd1 RhoGAP domain using a high yield expression procedure. By gel filtration and circular dichroïsm we provide the first evidences for a specific interaction between a RhoGAP domain (the RhoGAP domain of Rgd1p) and phosphoinositides.
authors.affiliation: Université de Bordeaux, Chimie et Biologie des Membranes et des Nanoobjets, CNRS UMR 5248, Pessac, France.
authors.forename: Benoît
authors.initials: B
authors.lastname: Odaert
authors.forename: Valérie
authors.initials: V
authors.lastname: Prouzet-Mauleon
authors.forename: Jean-William
authors.initials: JW
authors.lastname: Dupuy
authors.forename: Marc
authors.initials: M
authors.lastname: Crouzet
authors.forename: Marc
authors.initials: M
authors.lastname: Bonneu
authors.forename: Xavier
authors.initials: X
authors.lastname: Santarelli
authors.forename: Aurélie
authors.initials: A
authors.lastname: Vieillemard
authors.forename: Didier
authors.initials: D
authors.lastname: Thoraval
authors.forename: François
authors.initials: F
authors.lastname: Doignon
authors.forename: Michel
authors.initials: M
authors.lastname: Hugues
mesh_terms: D020690:GTPase-Activating Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010716:Phosphatidylinositols / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D020690:GTPase-Activating Proteins; D010716:Phosphatidylinositols; C449479:RGD1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C085379:rho GTPase-activating protein