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Related Experiment Videos

Promiscuity in Rab-SNARE interactions.

E Grote1, P J Novick

  • 1Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06520-8002, USA.

Molecular Biology of the Cell
|December 10, 1999
PubMed
Summary
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This study investigated if Rab proteins and SNAREs ensure yeast secretory vesicles fuse only with the plasma membrane. Results show Rab-Sec4p and SNARE interactions are nonselective, indicating they do not specify target membranes.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Yeast Genetics

Background:

  • Exocytosis in yeast involves Rab proteins and SNAREs for vesicle fusion.
  • Sec4p, a Rab protein, and SNAREs (Snc, Sso, Sec9) mediate post-Golgi vesicle fusion with the plasma membrane.
  • The mechanism ensuring vesicles fuse with the plasma membrane and not other organelles is unclear.

Purpose of the Study:

  • To test if selective interactions between Rab protein Sec4p and exocytic SNAREs ensure target membrane specificity.
  • To investigate the role of Rab-SNARE interactions in directing secretory vesicles to the plasma membrane.

Main Methods:

  • Analysis of SNARE complex assembly in a sec4-8 mutant yeast strain.
  • In vivo binding assays to study interactions between Sec4p and various t-SNAREs.

Related Experiment Videos

  • Co-immunoprecipitation assays using nucleotide-free forms of Rab proteins and SNARE proteins.

    • Main Results:

    • SNARE complex assembly is defective in sec4-8 mutants.
    • Sec4p binding to the endosomal/Golgi t-SNARE Tlg2p is reduced in sec4-8 cells.
    • Nucleotide-free Rab proteins, including Sec4p, bind nonselectively to multiple SNARE proteins (Ssop, Pep12p, Sncp).

    Conclusions:

    • Rab-SNARE interactions are not selective for the plasma membrane.
    • Rabs and SNAREs do not cooperate to specify the target membrane during yeast exocytosis.
    • The study challenges the hypothesis of selective Rab-SNARE interactions in directing vesicle fusion.