title: Rab GAP cascade regulates dynamics of Ypt6 in the Golgi traffic.
abstract: The Golgi apparatus functions as the central station of membrane traffic in cells, where newly synthesized proteins moving along the secretory pathway merge with proteins recycled from subsequent membrane organelles such as endosomes. A series of Rab GTPases act consecutively and in concert with the maturation of cis- to-trans cisternae of the Golgi apparatus. Rab GTPases control various steps in intracellular membrane traffic by recruiting downstream effector proteins. Here, we report the dynamics of Ypt6, a yeast member of the Rab GTPase family, which mediates the fusion of vesicles from endosomes at the Golgi apparatus. Ypt6 resides temporarily at the Golgi and dissociates into the cytosol upon arrival of Ypt32, another Rab GTPase functioning in the late Golgi. We found that Gyp6, a putative GTPase-activating protein (GAP) for Ypt6, specifically interacts with Ypt32, most likely as an effector. Disruption of GYP6 or introduction of a Rab-GAP activity-deficient mutation in GYP6 resulted in continual residence of Ypt6 at the Golgi. We propose that Ypt32 acts to terminate endosome-to-Golgi traffic through a Rab-GAP cascade as it does for cis-to-trans intra-Golgi traffic. Simultaneous disruption of GAP for early-acting Rab proteins in the Golgi showed appreciable defects in post-Golgi trafficking, but did not significantly affect cell growth.
authors.affiliation: Live Cell Molecular Imaging Research Team, RIKEN Center for Advanced Photonics, Wako, Saitama 351-0198, Japan.
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Suda
authors.forename: Kazuo
authors.initials: K
authors.lastname: Kurokawa
authors.forename: Ryogo
authors.initials: R
authors.lastname: Hirata
authors.forename: Akihiko
authors.initials: A
authors.lastname: Nakano
mesh_terms: D020690:GTPase-Activating Proteins / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism*; D015151:Immunoblotting; D008856:Microscopy, Fluorescence; D020559:Monomeric GTP-Binding Proteins / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D021381:Protein Transport / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D020691:rab GTP-Binding Proteins / Q000378:metabolism
chemical_list: D020690:GTPase-Activating Proteins; C079894:GYP6 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C121633:YPT32 protein, S cerevisiae; D020559:Monomeric GTP-Binding Proteins; C499307:YPT6 protein, S cerevisiae; D020691:rab GTP-Binding Proteins