Retrograde transport from the yeast Golgi is mediated by two ARF GAP proteins with overlapping function

Pak Phi Poon, Dan Cassel, Anne Spang, Miriam Rotman, Elah Pick, Richard A. Singer, Gerald C. Johnston

Research output: Contribution to journalArticlepeer-review

Abstract

ARF proteins, which mediate vesicular transport, have little or no intrinsic GTPase activity. They rely on the actions of GTPase-activating proteins (GAPs) for their function. The in vitro GTPase activity of the Saccharomyces cerevisiae ARF proteins Arf1 and Arf2 is stimulated by the yeast Gcs1 protein, and in vivo genetic interactions between arf and gcs1 mutations implicate Gcs1 in vesicular transport. However, the Gcs1 protein is dispensable, indicating that additional ARF GAP proteins exist. We show that the structurally related protein Glo3, which is also dispensable, also exhibits ARF GAP activity. Genetic and in vitro approaches reveal that Glo3 and Gcs1 have an overlapping essential function at the endoplasmic reticulum (ER)-Golgi stage of vesicular transport. Mutant cells deficient for both ARF GAPs cannot proliferate, undergo a dramatic accumulation of ER and are defective for protein transport between ER and Golgi. The glo3Δ and gcs1Δ single mutations each interact with a sec21 mutation that affects a component of COPI, which mediates vesicular transport within the ER-Golgi shuttle, while increased dosage of the BET1, BOS1 and SEC22 genes encoding members of a v-SNARE family that functions within the ER-Golgi alleviates the effects of a glo3Δ mutation. An in vitro assay indicates that efficient retrieval from the Golgi to the ER requires these two proteins. These findings suggest that Glo3 and Gcs1 ARF GAPs mediate retrograde vesicular transport from the Golgi to the ER.

Original languageEnglish
Pages (from-to)555-564
Number of pages10
JournalEMBO Journal
Volume18
Issue number3
DOIs
StatePublished - 1 Feb 1999
Externally publishedYes

Keywords

  • ARF GAP
  • GTPase-activating protein
  • Gcs1
  • Glo3
  • Saccharomyces cerevisae
  • Vesicular transport

ASJC Scopus subject areas

  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology
  • Molecular Biology
  • General Neuroscience

Fingerprint

Dive into the research topics of 'Retrograde transport from the yeast Golgi is mediated by two ARF GAP proteins with overlapping function'. Together they form a unique fingerprint.

Cite this