Insulin rapidly induces the biosynthesis of elongation factor 2

Richard M Levenson, A. C. Nairn, P. J. Blackshear

Research output: Contribution to journalArticle

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Abstract

Insulin increases the rate of overall protein synthesis in many cells and tissues, while inducing the preferential expression of individual proteins. To identify and characterize such proteins, NIH 3T3 cells stably expressing more than 106 human insulin receptors per cell (HIR 3.5; Whittaker, J., Okamoto, A.K., Thys, R., Bell, G.I., Steiner., D.F., and Hofmann, C.A. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 5237-5241) were treated with insulin in the presence of [35S]methionine, and labeled proteins were separated using ultra-high resolution ('giant') two-dimensional gel electrophoresis. Overall protein synthesis was enhanced as much as 3-fold by insulin treatment; the synthesis of approximately 1% of the 2,500 proteins visible on the gel autoradiographs was further selectively increased. By using immunoblotting, immunoprecipitation and comigration assays, we identified one of the rapidly induced proteins (M(r) 96,000; pI 6.8) as eukaryotic elongation factor 2 (EF-2), a major component of the protein translation apparatus. Insulin induced the synthesis of EF-2 within 20 min of treatment, with a half-maximal dose of 10-11 M. It was synthesized as a precursor form that was processed to a more basic mature species within 30 min. Long term treatment with insulin led to accumulation of EF-2 within the cell and prevented the substantial decrease in EF-2 concentration that occurred during serum deprivation. Finally, we found that insulin induction of EF-2 occurred normally in the presence of the RNA-transcription inhibitor, actinomycin D. Thus, insulin rapidly induced the synthesis of EF-2 predominantly or exclusively at the level of mRNA translation.

Original languageEnglish (US)
Pages (from-to)11904-11911
Number of pages8
JournalJournal of Biological Chemistry
Volume264
Issue number20
StatePublished - 1989
Externally publishedYes

Fingerprint

Peptide Elongation Factor 2
Biosynthesis
Insulin
Proteins
Protein Biosynthesis
Gels
NIH 3T3 Cells
Electrophoresis, Gel, Two-Dimensional
Dactinomycin
Immunoprecipitation
Immunoblotting
Methionine
Transcription
Electrophoresis
Assays
RNA
Tissue
Messenger RNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

Insulin rapidly induces the biosynthesis of elongation factor 2. / Levenson, Richard M; Nairn, A. C.; Blackshear, P. J.

In: Journal of Biological Chemistry, Vol. 264, No. 20, 1989, p. 11904-11911.

Research output: Contribution to journalArticle

Levenson, RM, Nairn, AC & Blackshear, PJ 1989, 'Insulin rapidly induces the biosynthesis of elongation factor 2', Journal of Biological Chemistry, vol. 264, no. 20, pp. 11904-11911.
Levenson, Richard M ; Nairn, A. C. ; Blackshear, P. J. / Insulin rapidly induces the biosynthesis of elongation factor 2. In: Journal of Biological Chemistry. 1989 ; Vol. 264, No. 20. pp. 11904-11911.
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