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An essential role for src tyrosine kinase/shc adapter protein complexes in gβy subunit-mediated mitogen-activated protein kinase activation

Publication ,  Journal Article
Luttrell, LM; Hawes, BE; Van Biesen, T; Lultrell, DK; Lefkowit, RJ
Published in: Journal of Investigative Medicine
January 1, 1996

Several G protein-coupled recepiors which interact with pertussis toxin-sensitive heterotrimenc G proteins mediate Ras-dependent activation of mitogen-activated protein (MAP) kinases. The mechanism involves Gβysubunit-mediated increases in tyrosine phosphorylation of the She adapter protein, GRB2/She complex formation and recruitment of Ras guanine nucleotide exchange factor activity. We investigated the role of the ubiquitous nonreceptor tyrosine kinase c-Src, in activation of the MAP kinase pathway via endogenous G protein-coupled lysophosphatidic acid (LPA) receptors or transient expression of Gβy subunits in COS-7 cells. In vitro kinase assays of She immunoprecipitates following LPA stimulation demonstrated rapid, (ransient recruitment of tyrosine kinase activity into She immune complexes. Recruitment of lyrosine kinase aclivity was pertussis toxin-sensitive and mimicked by cellular expression of Gβy subunits. Immunoblots for coprecipitated proteins in she immunoprecipitates revealed a transient association of She and c-Src following LPA stimulation which coincided with increases in She-associated tyrosine kinase activity and she tyrosine phosphorylation. LPA stimulation or expression of Gβy subuntis resulted in c-Src activation as assessed by increased c-Src autophosphorylation. Overexpression of wild type or constitutively active mutant c-Src, but not kinase inactive mutant c-Src, lead to increased tyrosine kinase activity in She immunoprecipitates, increased She tyrosine phosphorylation and Shc/GRB2 complex formation. MAP kinase activation resulting from LPA receptor stimulation, expression of Gβy suhumts or expression of e-Src was sensitive to dominant negatives of mSos, Ras and Raf Coexpression of Csk, which inactivates Src family kinases by phosphoryfating the regulatory C-terminai lyrosine residue, inhibited LPA stimulation of She [yrosine phosphorylalion, Shc/GRB2 complex formation and MAP kinase activation. These data suggest that Gβy subunit-mediated formation of Shc/e-Sre complexes and cSrc kinase activation are eariy events m Ras-dependent activation of MAP kmasc via pertussis toxin sensitive G protein-coupled receptors.

Duke Scholars

Published In

Journal of Investigative Medicine

ISSN

1708-8267

Publication Date

January 1, 1996

Volume

44

Issue

3

Related Subject Headings

  • General Clinical Medicine
  • 3202 Clinical sciences
  • 1103 Clinical Sciences
 

Citation

APA
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ICMJE
MLA
NLM
Luttrell, L. M., Hawes, B. E., Van Biesen, T., Lultrell, D. K., & Lefkowit, R. J. (1996). An essential role for src tyrosine kinase/shc adapter protein complexes in gβy subunit-mediated mitogen-activated protein kinase activation. Journal of Investigative Medicine, 44(3).
Luttrell, L. M., B. E. Hawes, T. Van Biesen, D. K. Lultrell, and R. J. Lefkowit. “An essential role for src tyrosine kinase/shc adapter protein complexes in gβy subunit-mediated mitogen-activated protein kinase activation.” Journal of Investigative Medicine 44, no. 3 (January 1, 1996).
Luttrell LM, Hawes BE, Van Biesen T, Lultrell DK, Lefkowit RJ. An essential role for src tyrosine kinase/shc adapter protein complexes in gβy subunit-mediated mitogen-activated protein kinase activation. Journal of Investigative Medicine. 1996 Jan 1;44(3).
Luttrell LM, Hawes BE, Van Biesen T, Lultrell DK, Lefkowit RJ. An essential role for src tyrosine kinase/shc adapter protein complexes in gβy subunit-mediated mitogen-activated protein kinase activation. Journal of Investigative Medicine. 1996 Jan 1;44(3).

Published In

Journal of Investigative Medicine

ISSN

1708-8267

Publication Date

January 1, 1996

Volume

44

Issue

3

Related Subject Headings

  • General Clinical Medicine
  • 3202 Clinical sciences
  • 1103 Clinical Sciences