Activation of mitogen-activated protein kinase/extracellular signal-regulated kinase kinase by G protein and tyrosine kinase oncoproteins

Anne M. Gardner, Richard Vaillancourt, Gary L. Johnson

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103 Citations (Scopus)

Abstract

Mitogen-activated protein kinases (MAPKs) are rapidly phosphorylated and activated in response to a variety of extracellular stimuli in many different cell types. The kinases that activate MAPK, the MAPK/ ERK Kinases (MEKs), are also activated by phosphorylation. We have studied the influence of specific oncogenes on the regulation of MEK activity in NIH3T3 and Rat1a fibroblasts. We show that a similar MEK activity phosphorylates and activates MAPK in both growth factor-stimulated (epidermal growth factor and thrombin) and oncogene (gip2, v-src, and v-raf)-transfected cells. Gip2 and v-Src activated MEK-1 in transfected Rat 1a cells, whereas v-Raf activated MEK-1 in transfected NIH3T3 cells. These cell-selective differences in MEK activation parallel constitutive MAPK activation in these cell lines. Stable expression of the v-ras oncogene resulted in little constitutive MEK activation in either cell line, even though both were highly transformed. The growth factor and oncoprotein regulated MEK activity co-fractionated by Mono S chromatography with the 45-kDa MEK-1 protein. We further demonstrate in NIH3T3 and Rat 1a cells that Raf-1 is activated, as measured by its ability to phosphorylate MEK-1, in response to epidermal growth factor but not thrombin. Thus, the regulatory network of protein kinases that activate MAPK converges at MEK but diverges with the kinases that phosphorylate and activate MEK.

Original languageEnglish (US)
Pages (from-to)17896-17901
Number of pages6
JournalJournal of Biological Chemistry
Volume268
Issue number24
StatePublished - Aug 25 1993
Externally publishedYes

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Oncogene Proteins
Mitogen-Activated Protein Kinase Kinases
Extracellular Signal-Regulated MAP Kinases
Mitogen-Activated Protein Kinases
Protein-Tyrosine Kinases
Phosphotransferases
Chemical activation
MAP Kinase Kinase 1
Epidermal Growth Factor
Thrombin
Rats
Intercellular Signaling Peptides and Proteins
Cells
src Genes
MAP Kinase Kinase Kinases
Cell Line
Phosphorylation
ras Genes
Fibroblasts
Chromatography

ASJC Scopus subject areas

  • Biochemistry

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Activation of mitogen-activated protein kinase/extracellular signal-regulated kinase kinase by G protein and tyrosine kinase oncoproteins. / Gardner, Anne M.; Vaillancourt, Richard; Johnson, Gary L.

In: Journal of Biological Chemistry, Vol. 268, No. 24, 25.08.1993, p. 17896-17901.

Research output: Contribution to journalArticle

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