Interaction of the Saccharomyces cerevisiae cortical actin patch protein Rvs167p with proteins involved in ER to Golgi vesicle trafficking.
We have used affinity chromatography to identify two proteins that bind to the SH3 domain of the actin cytoskeleton protein Rvs167p: Gyp5p and Gyl1p. Gyp5p has been shown to be a GTPase activating protein (GAP) for Ypt1p, a Rab GTPase involved in ER to Golgi trafficking; Gyl1p is a protein that resembles Gyp5p and has recently been shown to colocalize with and belong to the same protein complex as Gyp5p. We show that Gyl1p and Gyp5p interact directly with each other, likely through their carboxy-terminal coiled-coil regions. In assays of GAP activity, Gyp5p had GAP activity toward Ypt1p and we found that this activity was stimulated by the addition of Gyl1p. Gyl1p had no GAP activity toward Ypt1p. Genetic experiments suggest a role for Gyp5p and Gyl1p in ER to Golgi trafficking, consistent with their biochemical role. Since Rvs167p has a previously characterized role in endocytosis and we have shown here that it interacts with proteins involved in Golgi vesicle trafficking, we suggest that Rvs167p may have a general role in vesicle trafficking.
Friesen
Helena
H
Department of Molecular and Medical Genetics, University of Toronto, Ontario, Canada.
Colwill
Karen
K
Robertson
Karen
K
Schub
Oliver
O
Andrews
Brenda
B
D000595:Amino Acid Sequence; D015153:Blotting, Western; D002845:Chromatography; D003599:Cytoskeleton / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004705:Endocytosis; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D020690:GTPase-Activating Proteins / Q000235:genetics / Q000378:metabolism*; D005838:Genotype; D006056:Golgi Apparatus / Q000378:metabolism*; D047468:Immunoprecipitation; D008024:Ligands; D008840:Microfilament Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D010641:Phenotype; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013997:Time Factors; D020691:rab GTP-Binding Proteins / Q000378:metabolism; D018909:src Homology Domains
D020690:GTPase-Activating Proteins; C504621:Gyl1 protein, S cerevisiae; C504620:Gyp5 protein, S cerevisiae; D008024:Ligands; D008840:Microfilament Proteins; C082282:RVS167 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D004247:DNA; C505432:APP1 protein, S cerevisiae; C060755:YPT1 protein, S cerevisiae; D020691:rab GTP-Binding Proteins