Evidence for the genetic interaction between the actin-binding protein Vrp1 and the RhoGAP Rgd1 mediated through Rho3p and Rho4p in Saccharomyces cerevisiae.
The non-essential RGD1 gene from Saccharomyces cerevisiae encodes a protein that has been characterized in vitro as a Rho GTPase activating protein (RhoGAP) for the Rho3 and Rho4 proteins. Rgd1p, which displays a conserved FCH-coiled coil-Rho-GAP domain organization, showed a patch-like distribution in the cell, including a localization in growing buds. Using a genetic screen, we found that rgd1delta and vrp1alpha mutations exhibited a synthetic lethality, thus revealing an interaction between these genes. The VRP1 product is an actin and myosin interacting protein involved in polarized growth. Using mutant forms of both Rho3 and Rho4 proteins, we provide evidence for the involvement of these two GTPases in RGD1-VRP1 co-lethality. In addition, these results strongly argue in favour of Rho3p and Rho4p being the targets of Rgd1p RhoGAP activity in vivo. Genetic relationships between either VRP1 or RGD1 and actin cytoskeleton-linked genes were also studied. These and other well-established data support the idea that Vrp1, Las17, Rvs167 proteins belong to the same complex. This protein structure might act with myosins in various actin cytoskeleton-based activities, in co-operation with a Rho3p/Rho4p signalling pathway that is negatively regulated by Rgd1p GAP activity.
Roumanie
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Laboratoire de Biologie Moléculaire et de Séquençage, UMR CNRS 5095, Bordeaux, France.
Peypouquet
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MF
Bonneu
M
M
Thoraval
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D
Doignon
F
F
Crouzet
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M
D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D019204:GTP-Binding Proteins; D020690:GTPase-Activating Proteins / Q000235:genetics* / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D008840:Microfilament Proteins / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D020741:rho GTP-Binding Proteins / Q000235:genetics* / Q000378:metabolism
D005656:Fungal Proteins; D020690:GTPase-Activating Proteins; D008840:Microfilament Proteins; D029701:Saccharomyces cerevisiae Proteins; C090350:VRP1 protein, S cerevisiae; C085379:rho GTPase-activating protein; D019204:GTP-Binding Proteins; C074647:RHO3 protein, S cerevisiae; C074648:RHO4 protein, S cerevisiae; D020741:rho GTP-Binding Proteins