Phosphorylation of elongation factor 1 and ribosomal protein S6 by multipotential S6 kinase and insulin stimulation of translational elongation.
Stimulation of protein synthesis in response to insulin is concomitant with increased phosphorylation of initiation factors 4B and 4G and ribosomal protein S6 (Morley, S. J., and Traugh, J. A. (1993) Biochimie 75, 985-989) and is due at least in part to multipotential S6 kinase. When elongation factor 1 (EF-1) from rabbit reticulocytes was examined as substrate for multipotential S6 kinase, up to 1 mol/mol of phosphate was incorporated into the alpha, beta, and delta subunits. Phosphorylation of EF-1 resulted in a 2-2. 6-fold stimulation of EF-1 activity, as measured by poly(U)-directed polyphenylalanine synthesis. The rate of elongation was also stimulated by approximately 2-fold with 80 S ribosomes phosphorylated on S6 by multipotential S6 kinase. When the rates of elongation in extracts from serum-fed 3T3-L1 cells and cells serum-deprived for 1.5 h were compared, a 40% decrease was observed upon serum deprivation. The addition of insulin to serum-deprived cells for 15 min stimulated elongation to a rate equivalent to that of serum-fed cells. Similar results were obtained with partially purified EF-1, with both EF-1 and ribosomes contributing to stimulation of elongation. These data are consistent with a ribosomal transit time of 3.2 min for serum-deprived cells and 1.6 min following the addition of insulin for 15 min. Taken together, the data suggest that insulin stimulation involves coordinate regulation of EF-1 and ribosomes through phosphorylation by multipotential S6 kinase.
Chang
Y W
YW
Department of Biochemistry, University of California, Riverside, California 92521, USA.
Traugh
J A
JA
D016475:3T3 Cells; D000818:Animals; D007328:Insulin / Q000494:pharmacology; D051379:Mice; D010441:Peptide Chain Elongation, Translational; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors / Q000378:metabolism*; D010449:Peptide Mapping; D010766:Phosphorylation; D011817:Rabbits; D038601:Ribosomal Protein S6; D019893:Ribosomal Protein S6 Kinases / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000378:metabolism*
D007328:Insulin; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors; D038601:Ribosomal Protein S6; D012269:Ribosomal Proteins; D019893:Ribosomal Protein S6 Kinases