Evidence that phospholipase D mediates ADP ribosylation factor-dependent formation of Golgi coated vesicles

Nicholas T. Ktistakis, H. Alex Brown, M. Gerard Waters, Paul C. Sternweis, Michael G. Roth

Research output: Contribution to journalArticlepeer-review

329 Scopus citations


Formation of coatomer-coated vesicles from Golgi-enriched membranes requires the activation of a small GTP-binding protein, ADP ribosylation factor (ARF). ARF is also an efficacious activator of phospholipase D (PLD), an activity that is relatively abundant on Golgi-enriched membranes. It has been proposed that ARF, which is recruited onto membranes from cytosolic pools, acts directly to promote coatomer binding and is in a 3:1 stoichiometry with coatomer on coated vesicles. We present evidence that cytosolic ARF is not necessary for initiating coat assembly on Golgi membranes from cell lines with high constitutive PLD activity. Conditions are also described under which ARF is at most a minor component relative to coatomer in coated vesicles from all cell lines tested, including Chinese hamster ovary cells. Formation of coated vesicles was sensitive to ethanol at concentrations that inhibit the production of phosphatidic acid (PA) by PLD. When PA was produced in Golgi membranes by an exogenous bacterial PLD, rather than with ARF and endogenous PLD, coatomer bound to Golgi membranes. Purified coatomer also bound selectively to artificial lipid vesicles that contained PA and phosphatidylinositol (4,5)-bisphosphate (PIP2). We propose that activation of PLD and the subsequent production of PA are key early events for the formation of coatomer-coated vesicles.

Original languageEnglish (US)
Pages (from-to)295-306
Number of pages12
JournalJournal of Cell Biology
Issue number2
StatePublished - Jul 1996

ASJC Scopus subject areas

  • Cell Biology


Dive into the research topics of 'Evidence that phospholipase D mediates ADP ribosylation factor-dependent formation of Golgi coated vesicles'. Together they form a unique fingerprint.

Cite this