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The phosphatidylinositol 3-phosphate binding protein Vac1p interacts with a Rab GTPase and a Sec1p homologue to facilitate vesicle-mediated vacuolar protein sorting.

Activated GTP-bound Rab proteins are thought to interact with effectors to elicit vesicle targeting and fusion events. Vesicle-associated v-SNARE and target membrane t-SNARE proteins are also involved in vesicular transport. Little is known about the functional relationship between Rabs and SNARE protein complexes. We have constructed an activated allele of VPS21, a yeast Rab protein involved in vacuolar protein sorting, and demonstrated an allele-specific interaction between Vps21p and Vac1p. Vac1p was found to bind the Sec1p homologue Vps45p. Although no association between Vps21p and Vps45p was seen, a genetic interaction between VPS21 and VPS45 was observed. Vac1p contains a zinc-binding FYVE finger that may bind phosphatidylinositol 3-phosphate [PtdIns(3)P]. In other FYVE domain proteins, this motif and PtdIns(3)P are necessary for membrane association. Vac1 proteins with mutant FYVE fingers still associated with membranes but showed vacuolar protein sorting defects and reduced interactions with Vps45p and activated Vps21p. Vac1p membrane association was not dependent on PtdIns(3)P, Pep12p, Vps21p, Vps45p, or the PtdIns 3-kinase, Vps34p. Vac1p FYVE finger mutant missorting phenotypes were suppressed by a defective allele of VPS34. These data indicate that PtdIns(3)P may perform a regulatory role, possibly involved in mediating Vac1p protein-protein interactions. We propose that activated-Vps21p interacts with its effector, Vac1p, which interacts with Vps45p to regulate the Golgi to endosome SNARE complex.

Pubmed ID: 10359603

Authors

  • Tall GG
  • Hama H
  • DeWald DB
  • Horazdovsky BF

Journal

Molecular biology of the cell

Publication Data

June 29, 1999

Associated Grants

  • Agency: NIGMS NIH HHS, Id: GM-55301

Mesh Terms

  • Adaptor Proteins, Signal Transducing
  • Carboxypeptidases
  • Cathepsin A
  • Cell Membrane
  • Coated Vesicles
  • Cytoskeletal Proteins
  • Fungal Proteins
  • GTP-Binding Proteins
  • Hybrid Cells
  • Membrane Proteins
  • Munc18 Proteins
  • Nerve Tissue Proteins
  • Phosphatidylinositol 3-Kinases
  • Point Mutation
  • SNARE Proteins
  • Saccharomyces cerevisiae Proteins
  • Suppression, Genetic
  • Vacuoles
  • Vesicular Transport Proteins
  • Yeasts
  • Zinc Fingers
  • rab GTP-Binding Proteins
  • rab5 GTP-Binding Proteins