ADP-ribosylation Factor Proteins Mediate Agonist-induced Activation of Phospholipase D*

The role of small G proteins of the ADP-ribosylation factor (ARF) and Rho families on the activation of phospholipase D (PLD) by platelet-derived growth factor (PDGF) and phorbol esters (PMA) has been investigated. The activation of PLD by PDGF and PMA was blocked by brefeldin A (BFA), an inhibitor of ARF activation, but not by Clostridium botulinum C3 exotoxin, an inhibitor of the activity of Rho. PDGF and PMA, in the presence of GTPγS, promoted the association of ARF and RhoA with cell membranes. Cells depleted of ARF and Rho by digitonin permeabilization showed a significant reduction of the activity of phospholipase D. Recombinant ARF was sufficient to restore agonist-induced PLD activity to digitonin-permeabilized, cytoplasm-depleted cells. In contrast, isoprenylated recombinant RhoA had no effects in this reconstitution assay. HIRcB cells were transiently transfected with wild-type and dominant-negative mutants of ARF1 and ARF6. Neither wt-ARF1 nor wt-ARF6 had any effects on agonist-dependent PLD activity. However, dominant-negative ARF1 and ARF6 mutants blocked the stimulation of PLD by PDGF but only partially inhibited the effects of PMA. These results demonstrate that ARF rather than Rho proteins mediate the activation of PLD by PDGF and phorbol esters in HIRcB fibroblasts.

• Publication year

  1998

• Venue

  Journal of Biological Chemistry

• Publication date

  1998-11-13

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1074/jbc.273.46.30836PMID 9804862
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• ARF proteins rather than Rho proteins mediate agonist-induced activation of phospholipase D by PDGF and phorbol esters in HIRcB fibroblasts.

  Confidence 0.99

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• In HIRcB cells, wild-type ARF1 and ARF6 did not alter agonist-dependent PLD activity, but dominant-negative ARF1 and ARF6 mutants blocked PDGF-stimulated PLD and only partially inhibited PMA-stimulated PLD.

  Confidence 0.97

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• Digitonin permeabilization depleted ARF and Rho from cells and significantly reduced phospholipase D activity, and recombinant ARF restored agonist-induced PLD activity in the depleted cells.

  Confidence 0.96

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• Brefeldin A blocked PLD activation by PDGF and phorbol esters.

  Confidence 0.98

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review

• adp-ribosylation factor proteins

  protein family

  A family of small GTP-binding proteins examined here for their role in agonist-induced phospholipase D activation.

  Aliases: ARF proteins, ARF

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• arf1

  protein

  A specific ARF-family small GTPase tested as wild-type and dominant-negative constructs.

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• arf6

  protein

  A specific ARF-family small GTPase tested as wild-type and dominant-negative constructs.

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• brefeldin a

  inhibitor, small molecule

  An inhibitor used here to block ARF activation.

  Aliases: BFA

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• digitonin permeabilization

  experimental method

  A membrane-permeabilization procedure used to deplete cells of cytoplasmic factors before reconstitution.

  Aliases: digitonin-permeabilized cells

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• hircb cells

  cell line

  The fibroblast cell line used for transfection and agonist-response experiments.

  Aliases: HIRcB fibroblasts

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• phorbol esters

  agonist

  Protein kinase C-activating agonists represented in the experiments by PMA.

  Aliases: PMA

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• phospholipase d

  enzyme

  An enzyme whose activity was measured after agonist stimulation in the fibroblast assays.

  Aliases: PLD

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• platelet-derived growth factor

  growth factor

  A growth factor agonist used to stimulate phospholipase D in HIRcB fibroblasts.

  Aliases: PDGF

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• rho proteins

  protein family

  A family of small GTP-binding proteins compared with ARF proteins in the phospholipase D assays.

  Aliases: Rho family

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review

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