title: Yeast dynamin Vps1 and amphiphysin Rvs167 function together during endocytosis.
abstract: Dynamins are a conserved family of proteins involved in many membrane fusion and fission events. Previously, the dynamin-related protein Vps1 was shown to localize to endocytic sites, and yeast carrying deletions for genes encoding both the BAR domain protein Rvs167 and Vps1 had a more severe endocytic scission defect than either deletion alone. Vps1 and Rvs167 localize to endocytic sites at the onset of invagination and disassemble concomitant with inward vesicle movement. Rvs167-GFP localization is reduced in cells lacking vps1 suggesting that Vps1 influences Rvs167 association with the endocytic complex. Unlike classical dynamins, Vps1 does not have a proline-arginine domain that could interact with SH3 domain-containing proteins. Thus, while Rvs167 has an SH3 domain, it is not clear how an interaction would be mediated. Here, we demonstrate an interaction between Rvs167 SH3 domain and the single type I SH3-binding motif in Vps1. Mutant Vps1 that cannot bind Rvs167 rescues all membrane fusion/fission functions associated with Vps1 except for endocytic function, demonstrating the specificity and mechanistic importance of the interaction. In vitro, an Rvs161/Rvs167 heterodimer can disassemble Vps1 oligomers. Overall, the data support the idea that Vps1 and the amphiphysins function together to mediate scission during endocytosis in yeast.
authors.affiliation: Department of Molecular Biology and Biotechnology, Firth Court, Western Bank, University of Sheffield, Sheffield, S10 2TN, UK.
authors.forename: Iwona I
authors.initials: II
authors.lastname: Smaczynska-de Rooij
authors.forename: Ellen G
authors.initials: EG
authors.lastname: Allwood
authors.forename: Ritu
authors.initials: R
authors.lastname: Mishra
authors.forename: Wesley I
authors.initials: WI
authors.lastname: Booth
authors.forename: Soheil
authors.initials: S
authors.lastname: Aghamohammadzadeh
authors.forename: Martin W
authors.initials: MW
authors.lastname: Goldberg
authors.forename: Kathryn R
authors.initials: KR
authors.lastname: Ayscough
mesh_terms: D019943:Amino Acid Substitution / Q000502:physiology; D043402:Cathepsin A / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology*; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D008562:Membrane Glycoproteins / Q000378:metabolism; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D046912:Multiprotein Complexes / Q000378:metabolism; D011485:Protein Binding / Q000502:physiology; D054730:Protein Interaction Domains and Motifs / Q000502:physiology; D021381:Protein Transport / Q000502:physiology; D050683:R-SNARE Proteins / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion / Q000502:physiology; D020798:Two-Hybrid System Techniques; D014617:Vacuoles / Q000502:physiology; D033921:Vesicular Transport Proteins / Q000235:genetics / Q000378:metabolism*; D051316:Wiskott-Aldrich Syndrome Protein / Q000378:metabolism
chemical_list: D003598:Cytoskeletal Proteins; C480361:LAS17 protein, S cerevisiae; D008562:Membrane Glycoproteins; D008840:Microfilament Proteins; D046912:Multiprotein Complexes; D050683:R-SNARE Proteins; C089181:RVS161 protein, S cerevisiae; C082282:RVS167 protein, S cerevisiae; D011993:Recombinant Fusion Proteins; C082266:SLA2 protein, S cerevisiae; C083241:SNC1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D033921:Vesicular Transport Proteins; D051316:Wiskott-Aldrich Syndrome Protein; C026742:plastin; D043402:Cathepsin A; C499283:PRC1 protein, S cerevisiae; D019204:GTP-Binding Proteins; C064553:VPS1 protein, S cerevisiae