Ras/mitogen-activated protein kinase mediates norepinephrine-induced phospholipase D activation in rabbit aortic smooth muscle cells by a phosphorylation-dependent mechanism

Mubarack M. Muthalif, Jean Hugues Parmentier, Ibrahim F. Benter, Nour Karzoun, Aftab Ahmed, Zinat Khandekar, Mohamed Z. Adl, Sylvain Bourgoin, Kafait Malik

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Phospholipase D (PLD) activity is regulated by phosphatidylinositol 4,5- biphosphate, protein kinase C (PKC), ADP-ribosylation factor, and Rho. The present study was designed to investigate the mechanism of norepinephrine (NE)-mediated PLD activation in rabbit aortic vascular smooth muscle cells (VSMC). NE (10 μM) caused activation of PLD, as measured by the production of phosphatidylethanol in [3H]oleic acid-labeled cells. NE also increased PKC activity in VSMC. However, treatment of cells with bisindolylmaleimide, a PKC inhibitor, or long-term treatment with phorbol-12-myristate-13-acetate that depletes PKC did not decrease NE-induced activation of PLD. NE- stimulated PLD activity was attenuated by farnesyl transferase inhibitors (FPT III and SCH-56582), which reduce activation of both Ras and mitogen- activated protein (MAP) kinase. Moreover, transfection of VSMC with a dominant negative Ras resulted in inhibition of NE-stimulated MAP kinase and PLD activities. Treatment of cells with PD-98059, a MAP kinase kinase inhibitor, also reduced NE-stimulated PLD activity. These data suggest that NE-stimulated PLD activity is mediated via activation of Ras and MAP kinase in rabbit VSMC. To study the mechanism of activation of PLD by Ras/MAP kinase, NE-induced phosphorylation of PLD was examined. In VSMC, PLD of molecular mass 120 kDa was identified with polyclonal PLD antibody. Phosphorylation of PLD by NE, measured as 32P incorporation into PLD, was inhibited by PD-98059. Moreover, PLD immunoprecipitated from VSMC lysates was phosphorylated in vitro by MAP kinase. Collectively, these results show a novel pathway for activation of PLD that appears to be mediated through Ras/MAP kinase pathway by a mechanism involving phosphorylation.

Original languageEnglish (US)
Pages (from-to)268-274
Number of pages7
JournalJournal of Pharmacology and Experimental Therapeutics
Volume293
Issue number1
StatePublished - Apr 1 2000

Fingerprint

Phospholipase D
Mitogen-Activated Protein Kinases
Smooth Muscle Myocytes
Norepinephrine
Phosphorylation
Rabbits
Vascular Smooth Muscle
Protein Kinase C
Phosphatidylinositol 4,5-Diphosphate
ADP-Ribosylation Factors
Protein C Inhibitor
Mitogen-Activated Protein Kinase Kinases
Oleic Acid
Protein Kinase Inhibitors
Transferases

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Pharmacology

Cite this

Ras/mitogen-activated protein kinase mediates norepinephrine-induced phospholipase D activation in rabbit aortic smooth muscle cells by a phosphorylation-dependent mechanism. / Muthalif, Mubarack M.; Parmentier, Jean Hugues; Benter, Ibrahim F.; Karzoun, Nour; Ahmed, Aftab; Khandekar, Zinat; Adl, Mohamed Z.; Bourgoin, Sylvain; Malik, Kafait.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 293, No. 1, 01.04.2000, p. 268-274.

Research output: Contribution to journalArticle

Muthalif, Mubarack M. ; Parmentier, Jean Hugues ; Benter, Ibrahim F. ; Karzoun, Nour ; Ahmed, Aftab ; Khandekar, Zinat ; Adl, Mohamed Z. ; Bourgoin, Sylvain ; Malik, Kafait. / Ras/mitogen-activated protein kinase mediates norepinephrine-induced phospholipase D activation in rabbit aortic smooth muscle cells by a phosphorylation-dependent mechanism. In: Journal of Pharmacology and Experimental Therapeutics. 2000 ; Vol. 293, No. 1. pp. 268-274.
@article{73248cf0097d47deb1067db84edc3133,
title = "Ras/mitogen-activated protein kinase mediates norepinephrine-induced phospholipase D activation in rabbit aortic smooth muscle cells by a phosphorylation-dependent mechanism",
abstract = "Phospholipase D (PLD) activity is regulated by phosphatidylinositol 4,5- biphosphate, protein kinase C (PKC), ADP-ribosylation factor, and Rho. The present study was designed to investigate the mechanism of norepinephrine (NE)-mediated PLD activation in rabbit aortic vascular smooth muscle cells (VSMC). NE (10 μM) caused activation of PLD, as measured by the production of phosphatidylethanol in [3H]oleic acid-labeled cells. NE also increased PKC activity in VSMC. However, treatment of cells with bisindolylmaleimide, a PKC inhibitor, or long-term treatment with phorbol-12-myristate-13-acetate that depletes PKC did not decrease NE-induced activation of PLD. NE- stimulated PLD activity was attenuated by farnesyl transferase inhibitors (FPT III and SCH-56582), which reduce activation of both Ras and mitogen- activated protein (MAP) kinase. Moreover, transfection of VSMC with a dominant negative Ras resulted in inhibition of NE-stimulated MAP kinase and PLD activities. Treatment of cells with PD-98059, a MAP kinase kinase inhibitor, also reduced NE-stimulated PLD activity. These data suggest that NE-stimulated PLD activity is mediated via activation of Ras and MAP kinase in rabbit VSMC. To study the mechanism of activation of PLD by Ras/MAP kinase, NE-induced phosphorylation of PLD was examined. In VSMC, PLD of molecular mass 120 kDa was identified with polyclonal PLD antibody. Phosphorylation of PLD by NE, measured as 32P incorporation into PLD, was inhibited by PD-98059. Moreover, PLD immunoprecipitated from VSMC lysates was phosphorylated in vitro by MAP kinase. Collectively, these results show a novel pathway for activation of PLD that appears to be mediated through Ras/MAP kinase pathway by a mechanism involving phosphorylation.",
author = "Muthalif, {Mubarack M.} and Parmentier, {Jean Hugues} and Benter, {Ibrahim F.} and Nour Karzoun and Aftab Ahmed and Zinat Khandekar and Adl, {Mohamed Z.} and Sylvain Bourgoin and Kafait Malik",
year = "2000",
month = "4",
day = "1",
language = "English (US)",
volume = "293",
pages = "268--274",
journal = "Journal of Pharmacology and Experimental Therapeutics",
issn = "0022-3565",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "1",

}

TY - JOUR

T1 - Ras/mitogen-activated protein kinase mediates norepinephrine-induced phospholipase D activation in rabbit aortic smooth muscle cells by a phosphorylation-dependent mechanism

AU - Muthalif, Mubarack M.

AU - Parmentier, Jean Hugues

AU - Benter, Ibrahim F.

AU - Karzoun, Nour

AU - Ahmed, Aftab

AU - Khandekar, Zinat

AU - Adl, Mohamed Z.

AU - Bourgoin, Sylvain

AU - Malik, Kafait

PY - 2000/4/1

Y1 - 2000/4/1

N2 - Phospholipase D (PLD) activity is regulated by phosphatidylinositol 4,5- biphosphate, protein kinase C (PKC), ADP-ribosylation factor, and Rho. The present study was designed to investigate the mechanism of norepinephrine (NE)-mediated PLD activation in rabbit aortic vascular smooth muscle cells (VSMC). NE (10 μM) caused activation of PLD, as measured by the production of phosphatidylethanol in [3H]oleic acid-labeled cells. NE also increased PKC activity in VSMC. However, treatment of cells with bisindolylmaleimide, a PKC inhibitor, or long-term treatment with phorbol-12-myristate-13-acetate that depletes PKC did not decrease NE-induced activation of PLD. NE- stimulated PLD activity was attenuated by farnesyl transferase inhibitors (FPT III and SCH-56582), which reduce activation of both Ras and mitogen- activated protein (MAP) kinase. Moreover, transfection of VSMC with a dominant negative Ras resulted in inhibition of NE-stimulated MAP kinase and PLD activities. Treatment of cells with PD-98059, a MAP kinase kinase inhibitor, also reduced NE-stimulated PLD activity. These data suggest that NE-stimulated PLD activity is mediated via activation of Ras and MAP kinase in rabbit VSMC. To study the mechanism of activation of PLD by Ras/MAP kinase, NE-induced phosphorylation of PLD was examined. In VSMC, PLD of molecular mass 120 kDa was identified with polyclonal PLD antibody. Phosphorylation of PLD by NE, measured as 32P incorporation into PLD, was inhibited by PD-98059. Moreover, PLD immunoprecipitated from VSMC lysates was phosphorylated in vitro by MAP kinase. Collectively, these results show a novel pathway for activation of PLD that appears to be mediated through Ras/MAP kinase pathway by a mechanism involving phosphorylation.

AB - Phospholipase D (PLD) activity is regulated by phosphatidylinositol 4,5- biphosphate, protein kinase C (PKC), ADP-ribosylation factor, and Rho. The present study was designed to investigate the mechanism of norepinephrine (NE)-mediated PLD activation in rabbit aortic vascular smooth muscle cells (VSMC). NE (10 μM) caused activation of PLD, as measured by the production of phosphatidylethanol in [3H]oleic acid-labeled cells. NE also increased PKC activity in VSMC. However, treatment of cells with bisindolylmaleimide, a PKC inhibitor, or long-term treatment with phorbol-12-myristate-13-acetate that depletes PKC did not decrease NE-induced activation of PLD. NE- stimulated PLD activity was attenuated by farnesyl transferase inhibitors (FPT III and SCH-56582), which reduce activation of both Ras and mitogen- activated protein (MAP) kinase. Moreover, transfection of VSMC with a dominant negative Ras resulted in inhibition of NE-stimulated MAP kinase and PLD activities. Treatment of cells with PD-98059, a MAP kinase kinase inhibitor, also reduced NE-stimulated PLD activity. These data suggest that NE-stimulated PLD activity is mediated via activation of Ras and MAP kinase in rabbit VSMC. To study the mechanism of activation of PLD by Ras/MAP kinase, NE-induced phosphorylation of PLD was examined. In VSMC, PLD of molecular mass 120 kDa was identified with polyclonal PLD antibody. Phosphorylation of PLD by NE, measured as 32P incorporation into PLD, was inhibited by PD-98059. Moreover, PLD immunoprecipitated from VSMC lysates was phosphorylated in vitro by MAP kinase. Collectively, these results show a novel pathway for activation of PLD that appears to be mediated through Ras/MAP kinase pathway by a mechanism involving phosphorylation.

UR - http://www.scopus.com/inward/record.url?scp=0034038211&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034038211&partnerID=8YFLogxK

M3 - Article

VL - 293

SP - 268

EP - 274

JO - Journal of Pharmacology and Experimental Therapeutics

JF - Journal of Pharmacology and Experimental Therapeutics

SN - 0022-3565

IS - 1

ER -