Phosphorylation of elongation factor 1 (EF-1) by protein kinase C stimulates GDP/GTP-exchange activity.
Phosphorylation of the alpha, beta and delta subunits of elongation factor (EF) 1 by protein kinase C results in stimulation of elongation activity up to threefold both in vivo and in vitro [Venema, R. C., Peters, H. I. & Traugh, J. A. (1991) J. Biol. Chem. 266, 11,993-11,998, Venema, R. C., Peters, H. I. & Traugh, J. A. (1991) J. Biol. Chem. 266, 12,574-12,580]. The alpha subunit catalyzes the GTP-dependent binding of amino-acyl-tRNA to the ribosome, while the beta gamma and delta subunits of EF-1 catalyze exchange of the residual GDP on EF-1 alpha for GTP. To determine whether the change in elongation rate following phosphorylation by protein kinase C is due to stimulation of GDP/GTP exchange activity, EF-1 and EF-1.valyl-tRNA-synthetase have been purified from rabbit reticulocytes, phosphorylated in vitro by protein kinase C and the effect of phosphorylation on nucleotide-exchange activity analyzed. The alpha, beta and delta subunits are phosphorylated only on serine, and phosphopeptide maps show distinct phosphopeptides for each subunit. Following quantitative phosphorylation of EF-1 by protein kinase C on the alpha, beta, and delta subunits, a twofold enhancement of the rate of nucleotide exchange over the non-phosphorylated controls is observed with EF-1 and EF-1.valyl-tRNA synthetase. Stimulation of nucleotide exchange results in a two-fold increase in the formation of EF-1 alpha.GTP.Phe-tRNA, leading to an increased rate of binding of Phe-tRNA to ribosomes. The magnitude of stimulation of the exchange rate is similar to that reported previously for the rate of elongation following phosphorylation of EF-1 by protein kinase C. Thus, the enhancement of EF-1 activity in response to 4 beta-phorbol 12-myristate 13-acetate appears to be due to stimulation of the rate of GDP/GTP exchange following phosphorylation of EF-1 by protein kinase C.
Department of Biochemistry, University of California, Riverside 92521, USA.
H I
HI
Peters
Y W
YW
Chang
J A
JA
Traugh
D000818:Animals; D006153:Guanosine Diphosphate / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000378:metabolism*; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors / Q000378:metabolism*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000494:pharmacology*; D011817:Rabbits; D014637:Valine-tRNA Ligase / Q000378:metabolism
D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors; D006153:Guanosine Diphosphate; D006160:Guanosine Triphosphate; D011493:Protein Kinase C; D014637:Valine-tRNA Ligase