Protein-protein Interactions of ESCRT Complexes in the Yeast Saccharomyces Cerevisiae

Traffic. 2004 Mar;5(3):194-210. doi: 10.1111/j.1600-0854.2004.00169.x.

Abstract

Ten class E Vps proteins in yeast are known components of the ESCRT complexes I, II and III, which are required for the sorting of proteins to the lumenal membranes of multivesicular bodies. We used the yeast 2 hybrid system to analyze the protein-protein interactions of all 17 soluble class E Vps proteins, as well as proteins thought to be required for the ubiquitination and deubiquitination of cargo proteins at multivesicular bodies. We identified novel interactions between yeast ESCRT complex components suggesting that ESCRTI binds to both ESCRTII and ESCRTIII. These interactions were confirmed by GST pull-down experiments. Our data indicate that the link between ESCRTI and ESCRTIII is via Vps28p and Vps37p/Srn2p binding directly to Vps20p, as well as through indirect interactions via ESCRTII. This is in contrast to the situation in mammalian cells where ESCRTI and ESCRTIII interact indirectly via ALIX, the mammalian homologue of yeast proteins Vps31p/Bro1p and Rim20p. Our data also enable us to link all soluble class E Vps proteins to the ESCRT complexes. We propose the formation of a large multimeric complex on the endosome membrane consisting of ESCRTI, ESCRTII, ESCRTIII and other associated proteins.

Publication types

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

MeSH terms

  • Adenosine Triphosphatases / genetics
  • Adenosine Triphosphatases / metabolism
  • Amino Acid Sequence
  • Calcium Chloride / pharmacology
  • Culture Media
  • Endopeptidases / metabolism
  • Endosomal Sorting Complexes Required for Transport
  • Genes, Fungal / genetics
  • Humans
  • Ions / metabolism
  • Ions / pharmacology
  • Lithium Chloride / pharmacology
  • Molecular Sequence Data
  • Multiprotein Complexes / chemistry
  • Multiprotein Complexes / genetics
  • Multiprotein Complexes / metabolism*
  • Mutation / genetics
  • Nuclear Proteins / metabolism
  • Protein Binding
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / growth & development
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Sequence Alignment
  • Two-Hybrid System Techniques
  • Ubiquitin Thiolesterase

Substances

  • Culture Media
  • DOA4 protein, S cerevisiae
  • Endosomal Sorting Complexes Required for Transport
  • Ions
  • Multiprotein Complexes
  • Nuclear Proteins
  • SNF7 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • VPS4 protein, S cerevisiae
  • Endopeptidases
  • Ubiquitin Thiolesterase
  • Adenosine Triphosphatases
  • Lithium Chloride
  • Calcium Chloride