Insulin stimulates the phosphorylation of the 66- and 52-kilodalton Shc isoforms by distinct pathways

Aimee W. Kao, Steven B. Waters, Shuichi Okada, Jeffrey E. Pessin

Research output: Contribution to journalArticle

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Abstract

In contrast to the 52-kDa Shc isoform, insulin stimulation caused a quantitative, time-dependent decrease in the SDS-PAGE mobility of 66-kDa Shc in both Chinese hamster ovary/IR cells and 3T3L1 adipocytes. Alkaline phosphatase treatment and direct phoamino acid analysis demonstrated that insulin stimulated an increase in serine phosphorylation of the 66-kDa isoform but not 52-kDa Shc, although the latter displayed a marked increase in tyrosine phosphorylation. To identify the responsible kinase pathway, we compared the effects on 66-kDa Shc serine phosphorylation by insulin, anisomycin, and osmotic shock, agents that specifically activate the ERK, JNK, or both pathways, respectively. Insulin and osmotic shock both stimulated a decrease in 66-kDa Shc mobility, whereas anisomycin had no effect. Furthermore, expression of a dominant-interfering Ras mutant (N17Ras) prevented the insulin-stimulated, but not the osmotic shock-induced serine phosphorylation of 66-kDa Shc Consistent with a MEK-dependent pathway mediating 66-kDa Shc serine phosphorylation, the specific MEK inhibitor (PD98059) and expression of a dominant-interfering MEK mutant partially inhibited both the insulin and osmotic shock-induced reduction in 66-kDa Shc mobility. In contrast, expression of the MAP kinase phosphatase (MKP-1) completely prevented ERK activation but did not inhibit the serine phosphorylation of 66-kDa Shc. These data demonstrate that insulin stimulates the serine phosphorylation of the 66-kDa Shc isoform through a MEK-dependent mechanism.

Original languageEnglish (US)
Pages (from-to)2474-2480
Number of pages7
JournalEndocrinology
Volume138
Issue number6
DOIs
StatePublished - 1997
Externally publishedYes

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Protein Isoforms
Serine
Phosphorylation
Insulin
Insulin Coma
Mitogen-Activated Protein Kinase Kinases
Osmotic Pressure
Anisomycin
Dual Specificity Phosphatase 1
Cricetulus
Adipocytes
Alkaline Phosphatase
Tyrosine
Polyacrylamide Gel Electrophoresis
Ovary
Phosphotransferases
Acids

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Insulin stimulates the phosphorylation of the 66- and 52-kilodalton Shc isoforms by distinct pathways. / Kao, Aimee W.; Waters, Steven B.; Okada, Shuichi; Pessin, Jeffrey E.

In: Endocrinology, Vol. 138, No. 6, 1997, p. 2474-2480.

Research output: Contribution to journalArticle

Kao, Aimee W. ; Waters, Steven B. ; Okada, Shuichi ; Pessin, Jeffrey E. / Insulin stimulates the phosphorylation of the 66- and 52-kilodalton Shc isoforms by distinct pathways. In: Endocrinology. 1997 ; Vol. 138, No. 6. pp. 2474-2480.
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