Reciprocal signaling between heterotrimeric G proteins and the p21- stimulated protein kinase

Jun Wang, Jeffrey A. Frost, Melanie H. Cobb, Elliott M. Ross

Research output: Contribution to journalArticle

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Abstract

p21-activated protein kinase (PAK)-1 phosphorylated Gα(z), a member of the Gα(i) family that is found in the brain, platelets, and adrenal medulla. Phosphorylation approached 1 mol of phosphate/mol of Gα(z) in vitro. In transfected cells, Gα(z) was phosphorylated both by wild-type PAK1 when stimulated by the GTP-binding protein Rac1 and by constitutively active PAK1 mutants. In vitro, phosphorylation occurred only at Ser16, one of two Ser residues that are the major substrate sites for protein kinase C (PKC). PAK1 did not phosphorylate other Gα subunits (i1, i2, i3, o, s, or q). PAK1- phosphorylated Gα(z) was resistant both to RGSZ1, a G(z)-selective GTPase- activating protein (GAP), and to RGS4, a relatively nonselective GAP for the G(i) and G(q) families of G proteins. Phosphorylation of Set27 by PKC did not alter sensitivity to either GAP. The previously described inhibition of G(z) GAPs by PKC is therefore mediated by phosphorylation of Ser16. Phosphorylation of either Ser16 by PAK1 or Ser27 by PKC decreased the affinity of Gα(z) for Gβγ; phosphorylation of both residues by PKC caused no further effect. PAK1 thus regulates Gα(z) function by attenuating the inhibitory effects of both GAPs and Gβγ. In this context, the kinase activity of PAK1 toward several protein substrates was directly inhibited by Gβγ, suggesting that PAK1 acts as a Gβγ-regulated effector protein. This inhibition of mammalian PAK1 by Gβγ contrasts with the stimulation of the PAK homolog Ste20p in Saccharomyces cerevisiae by the Gβγ homolog Ste4p/Ste18p.

Original languageEnglish (US)
Pages (from-to)31641-31647
Number of pages7
JournalJournal of Biological Chemistry
Volume274
Issue number44
DOIs
StatePublished - Oct 29 1999

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Heterotrimeric GTP-Binding Proteins
Phosphorylation
Protein Kinases
GTPase-Activating Proteins
Protein Kinase C
p21-Activated Kinases
rac1 GTP-Binding Protein
Gastrin-Secreting Cells
Adrenal Medulla
Substrates
Platelets
GTP-Binding Proteins
Yeast
Saccharomyces cerevisiae
Brain
Proteins
Phosphotransferases
Blood Platelets
Phosphates

ASJC Scopus subject areas

  • Biochemistry

Cite this

Reciprocal signaling between heterotrimeric G proteins and the p21- stimulated protein kinase. / Wang, Jun; Frost, Jeffrey A.; Cobb, Melanie H.; Ross, Elliott M.

In: Journal of Biological Chemistry, Vol. 274, No. 44, 29.10.1999, p. 31641-31647.

Research output: Contribution to journalArticle

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abstract = "p21-activated protein kinase (PAK)-1 phosphorylated Gα(z), a member of the Gα(i) family that is found in the brain, platelets, and adrenal medulla. Phosphorylation approached 1 mol of phosphate/mol of Gα(z) in vitro. In transfected cells, Gα(z) was phosphorylated both by wild-type PAK1 when stimulated by the GTP-binding protein Rac1 and by constitutively active PAK1 mutants. In vitro, phosphorylation occurred only at Ser16, one of two Ser residues that are the major substrate sites for protein kinase C (PKC). PAK1 did not phosphorylate other Gα subunits (i1, i2, i3, o, s, or q). PAK1- phosphorylated Gα(z) was resistant both to RGSZ1, a G(z)-selective GTPase- activating protein (GAP), and to RGS4, a relatively nonselective GAP for the G(i) and G(q) families of G proteins. Phosphorylation of Set27 by PKC did not alter sensitivity to either GAP. The previously described inhibition of G(z) GAPs by PKC is therefore mediated by phosphorylation of Ser16. Phosphorylation of either Ser16 by PAK1 or Ser27 by PKC decreased the affinity of Gα(z) for Gβγ; phosphorylation of both residues by PKC caused no further effect. PAK1 thus regulates Gα(z) function by attenuating the inhibitory effects of both GAPs and Gβγ. In this context, the kinase activity of PAK1 toward several protein substrates was directly inhibited by Gβγ, suggesting that PAK1 acts as a Gβγ-regulated effector protein. This inhibition of mammalian PAK1 by Gβγ contrasts with the stimulation of the PAK homolog Ste20p in Saccharomyces cerevisiae by the Gβγ homolog Ste4p/Ste18p.",
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AB - p21-activated protein kinase (PAK)-1 phosphorylated Gα(z), a member of the Gα(i) family that is found in the brain, platelets, and adrenal medulla. Phosphorylation approached 1 mol of phosphate/mol of Gα(z) in vitro. In transfected cells, Gα(z) was phosphorylated both by wild-type PAK1 when stimulated by the GTP-binding protein Rac1 and by constitutively active PAK1 mutants. In vitro, phosphorylation occurred only at Ser16, one of two Ser residues that are the major substrate sites for protein kinase C (PKC). PAK1 did not phosphorylate other Gα subunits (i1, i2, i3, o, s, or q). PAK1- phosphorylated Gα(z) was resistant both to RGSZ1, a G(z)-selective GTPase- activating protein (GAP), and to RGS4, a relatively nonselective GAP for the G(i) and G(q) families of G proteins. Phosphorylation of Set27 by PKC did not alter sensitivity to either GAP. The previously described inhibition of G(z) GAPs by PKC is therefore mediated by phosphorylation of Ser16. Phosphorylation of either Ser16 by PAK1 or Ser27 by PKC decreased the affinity of Gα(z) for Gβγ; phosphorylation of both residues by PKC caused no further effect. PAK1 thus regulates Gα(z) function by attenuating the inhibitory effects of both GAPs and Gβγ. In this context, the kinase activity of PAK1 toward several protein substrates was directly inhibited by Gβγ, suggesting that PAK1 acts as a Gβγ-regulated effector protein. This inhibition of mammalian PAK1 by Gβγ contrasts with the stimulation of the PAK homolog Ste20p in Saccharomyces cerevisiae by the Gβγ homolog Ste4p/Ste18p.

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