New component of the vacuolar class C-Vps complex couples nucleotide exchange on the Ypt7 GTPase to SNARE-dependent docking and fusion

J Cell Biol. 2000 Oct 30;151(3):551-62. doi: 10.1083/jcb.151.3.551.

Abstract

The class C subset of vacuolar protein sorting (Vps) proteins (Vps11, Vps18, Vps16 and Vps33) assembles into a vacuole/prevacuole-associated complex. Here we demonstrate that the class C-Vps complex contains two additional proteins, Vps39 and Vps41. The COOH-terminal 148 amino acids of Vps39 direct its association with the class C-Vps complex by binding to Vps11. A previous study has shown that a large protein complex containing Vps39 and Vps41 functions as a downstream effector of the active, GTP-bound form of Ypt7, a rab GTPase required for the fusion of vesicular intermediates with the vacuole (Price, A., D. Seals, W. Wickner, and C. Ungermann. 2000. J. Cell Biol. 148:1231-1238). Here we present data that indicate that this complex also functions to stimulate nucleotide exchange on Ypt7. We show that Vps39 directly binds the GDP-bound and nucleotide-free forms of Ypt7 and that purified Vps39 stimulates nucleotide exchange on Ypt7. We propose that the class C-Vps complex both promotes Vps39-dependent nucleotide exchange on Ypt7 and, based on the work of Price et al., acts as a Ypt7 effector that tethers transport vesicles to the vacuole. Thus, the class C-Vps complex directs multiple reactions during the docking and fusion of vesicles with the vacuole, each of which contributes to the overall specificity and efficiency of this transport process.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adaptor Proteins, Vesicular Transport
  • Amino Acid Sequence
  • Biological Transport
  • Carrier Proteins / metabolism
  • Conserved Sequence
  • Fungal Proteins / metabolism
  • Genes, Essential / genetics
  • Guanosine Triphosphate / metabolism*
  • Macromolecular Substances
  • Membrane Fusion*
  • Membrane Proteins / chemistry
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Models, Biological
  • Molecular Sequence Data
  • Mutation / genetics
  • Nuclear Proteins*
  • Protein Binding
  • Protein Structure, Tertiary
  • RNA-Binding Proteins / metabolism
  • SNARE Proteins
  • Saccharomyces cerevisiae / cytology
  • Saccharomyces cerevisiae / enzymology
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins*
  • Sequence Alignment
  • Substrate Specificity
  • Two-Hybrid System Techniques
  • Vacuoles / chemistry
  • Vacuoles / enzymology
  • Vacuoles / metabolism*
  • Vesicular Transport Proteins*
  • rab GTP-Binding Proteins / genetics
  • rab GTP-Binding Proteins / metabolism*

Substances

  • Adaptor Proteins, Vesicular Transport
  • Carrier Proteins
  • Fungal Proteins
  • Macromolecular Substances
  • Membrane Proteins
  • NPL3 protein, S cerevisiae
  • Nuclear Proteins
  • PEP5 protein, S cerevisiae
  • RNA-Binding Proteins
  • SNARE Proteins
  • Saccharomyces cerevisiae Proteins
  • VAM6 protein, S cerevisiae
  • VPS41 protein, S cerevisiae
  • Vesicular Transport Proteins
  • Guanosine Triphosphate
  • YPT7 protein, S cerevisiae
  • rab GTP-Binding Proteins