Phosphorylation of ribosomal protein S6 by cAMP-dependent protein kinase and mitogen-stimulated S6 kinase differentially alters translation of globin mRNA.
The effects of phosphorylation of ribosomal protein S6 by two different protein kinases, the cAMP-dependent protein kinase and the mitogen-stimulated S6 kinase, or translation of globin mRNA in a reconstituted system and on binding of globin mRNA to 40 S ribosomal subunits were examined. The cAMP-dependent protein kinase incorporated 1.5 mol of phosphate/mol of 40 S ribosomal subunits. Phosphorylation of S6 by the cAMP-dependent protein kinase had no effect on binding of 3' terminus-labeled globin mRNA to 40 S ribosomal subunits. [3H]Leucine incorporation with 40 S ribosomal subunits phosphorylated by the cAMP-dependent protein kinase was identical to that observed with nonphosphorylated 40 S ribosomal subunits, although on occasion, a slight inhibition (less than 10%) was observed; there was no effect on the rate of synthesis of either the alpha or beta chains of globin. Phosphorylation with the mitogen-stimulated S6 kinase (2.5 mol/mol) did not alter binding of globin mRNA to 40 S ribosomal subunits; however, translation of globin mRNA in the reconstituted protein-synthesizing system was stimulated up to 4-fold over that observed with nonphosphorylated subunits. Synthesis of both the alpha and beta chains of globin was enhanced by phosphorylation as shown by electrophoretic analysis. Since the sites phosphorylated by the mitogen-stimulated S6 kinase are identical to those observed in vivo in response to insulin and growth-promoting compounds, the data support the hypothesis that enhanced synthesis of specific proteins may be due to phosphorylation of S6 and that differential phosphorylation of S6 can alter translation of natural mRNA.
D000818:Animals; D005914:Globins / Q000235:genetics*; D046911:Macromolecular Substances; D008934:Mitogens / Q000494:pharmacology; D010766:Phosphorylation; D014176:Protein Biosynthesis; D011494:Protein Kinases / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics*; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism; D038601:Ribosomal Protein S6; D019893:Ribosomal Protein S6 Kinases; D012269:Ribosomal Proteins / Q000235:genetics / Q000378:metabolism*
D046911:Macromolecular Substances; D008934:Mitogens; D012333:RNA, Messenger; D038601:Ribosomal Protein S6; D012269:Ribosomal Proteins; D005914:Globins; D011494:Protein Kinases; D019893:Ribosomal Protein S6 Kinases