Phosphorylation of eukaryotic protein synthesis initiation factors.

Abstract
Phosphorylation of eukaryotic initiation factors was examined both in intact cells and in vitro with purified components. Intact rabbit reticulocytes were incubated in a medium containing [32P]phosphate, and 8 initiation factors were isolated and partially purified. The purified factors were analyzed on dodecyl sulfate/polyacrylamide gels and compared with highly purified nonradioactive factors. Significant amounts of radioactivity were found associated with initiation factors eIF-2, polypeptide 2 (MW 53,000); eIF-3, polypeptides 2 and 4 (MW 110,000 and 67,000); and eIF-4B. Purified initiation factors from rabbit reticulocytes were also treated in vitro with [.gamma.-32P]ATP and a cyclic[c]AMP-independent protein kinase isolated from rabbit erythrocytes. Only the factor polypeptides phosphorylated intracellularly were phosphorylated in vitro. The results suggest that the cAMP-independent protein kinase is responsible for the phosphorylation of specific initiation factors in cells active in protein synthesis and that it may play a role in regulating translation.

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