Expression and function of IRS-1 in insulin signal transmission

Xiao Jian Sun, Montserrat Miralpeix, Martin G. Myers, Erin M. Glasheen, Jonathan M. Backer, C. Ronald Kahn, Morris F. White

Research output: Contribution to journalArticle

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Abstract

IRS-1 is a major insulin receptor substrate which may play an important role in insulin signal transmission. The mRNA for IRS-1 in rat cells and tissues is about 9.5 kilobases (kb). Rat liver IRS-1 was stably expressed in Chinese hamster ovary (CHO) cells (CHO/ IRS-1). Although its calculated molecular mass is 131 kDa, IRS-1 from quiescent cells migrated between 165 and 170 kDa during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. IRS-1 was phosphorylated strongly on serine residues and weakly on threonine residues before insulin stimulation. Insulin immediately stimulated tyrosine phosphorylation of IRS-1, and after 10-30 min with insulin its apparent molecular mass increased to 175-180 kDa. Expression of the human insulin receptor and rat IRS-1 together in CHO/IR/IRS-1 cells increased the basal serine phosphorylation of IRS-1 and strongly increased tyrosine phosphorylation during insulin stimulation. Purified insulin receptors directly phosphorylated baculovirus-produced IRS-1 exclusively on tyrosine residues. By immunofluorescence, IRS-1 was absent from the nucleus, but otherwise distributed uniformly before and after insulin stimulation. Some IRS-1 associated with the insulin receptor during insulin stimulation. In addition, a phosphatidylinositol 3′-kinase associated with IRS-1 during insulin stimulation, and this association was more sensitive to insulin in CHO cells overexpressing the insulin receptor (CHO/IR cells), more responsive to insulin in CHO/IRS-1 cells, and both sensitive and responsive in CHO/IR/IRS-1 cells. Similarly, insulin-stimulated DNA synthesis was more sensitive to insulin in CHO/IR cells, and more responsive in CHO/IRS-1 cells; however, insulin-stimulated DNA synthesis was sensitive but poorly responsive to insulin in CHO/IR/IRS-1 cells. Together, these results suggest that IRS-1 is a direct physiologic substrate of the insulin receptor and may play an important role in insulin signal transmission.

Original languageEnglish (US)
Pages (from-to)22662-22672
Number of pages11
JournalJournal of Biological Chemistry
Volume267
Issue number31
StatePublished - Nov 5 1992
Externally publishedYes

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Insulin
Cricetulus
Ovary
Insulin Receptor
Phosphorylation
Tyrosine
Rats
Molecular mass
Serine
Phosphatidylinositol 3-Kinase
Baculoviridae
DNA
Substrates
Threonine
Electrophoresis
Sodium Dodecyl Sulfate
Liver
Fluorescent Antibody Technique
Polyacrylamide Gel Electrophoresis
Cells

ASJC Scopus subject areas

  • Biochemistry

Cite this

Sun, X. J., Miralpeix, M., Myers, M. G., Glasheen, E. M., Backer, J. M., Kahn, C. R., & White, M. F. (1992). Expression and function of IRS-1 in insulin signal transmission. Journal of Biological Chemistry, 267(31), 22662-22672.

Expression and function of IRS-1 in insulin signal transmission. / Sun, Xiao Jian; Miralpeix, Montserrat; Myers, Martin G.; Glasheen, Erin M.; Backer, Jonathan M.; Kahn, C. Ronald; White, Morris F.

In: Journal of Biological Chemistry, Vol. 267, No. 31, 05.11.1992, p. 22662-22672.

Research output: Contribution to journalArticle

Sun, XJ, Miralpeix, M, Myers, MG, Glasheen, EM, Backer, JM, Kahn, CR & White, MF 1992, 'Expression and function of IRS-1 in insulin signal transmission', Journal of Biological Chemistry, vol. 267, no. 31, pp. 22662-22672.
Sun XJ, Miralpeix M, Myers MG, Glasheen EM, Backer JM, Kahn CR et al. Expression and function of IRS-1 in insulin signal transmission. Journal of Biological Chemistry. 1992 Nov 5;267(31):22662-22672.
Sun, Xiao Jian ; Miralpeix, Montserrat ; Myers, Martin G. ; Glasheen, Erin M. ; Backer, Jonathan M. ; Kahn, C. Ronald ; White, Morris F. / Expression and function of IRS-1 in insulin signal transmission. In: Journal of Biological Chemistry. 1992 ; Vol. 267, No. 31. pp. 22662-22672.
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