Identification of a ribosomal protein S6 kinase regulated by transformation and growth-promoting stimuli.
A serine protein kinase specific for ribosomal protein S6 in 40 S subunits has been identified and purified greater than 15,000-fold (with 18% recovery) from developing chicken embryos. An analogous enzyme has also been detected in serum-stimulated chicken embryo fibroblasts. The S6 kinase was identified as a phosphoprotein of Mr approximately 65,000 based on (i) gel filtration, (ii) apparent autophosphorylation of a 65-kDa protein when several enzyme preparations were incubated with [gamma-32P]ATP in the absence of added substrate, (iii) comigration of S6 kinase activity with the autophosphorylating activity over a variety of chromatographic resins, and (iv) elution and renaturation of S6 kinase activity from the 65-kDa region of a sodium dodecyl sulfate-polyacrylamide gel. The purified protein kinase is highly specific for S6 in 40 S subunits and does not appreciably phosphorylate casein, histone H1, mixed histones, protamine, polyoma virus capsid protein, or phosphorylase a/b. These characteristics suggest that this enzyme is unrelated to other protein kinases believed to be activated in stimulated cells, including cAMP-dependent protein kinase, protein kinase C (Ca2+/phospholipid-dependent enzyme), or Ca2+/calmodulin-dependent protein kinases. In fibroblasts, S6 kinase is activated by a variety of mitogenic agents including the tyrosine-specific protein kinase of Rous sarcoma virus, pp60v-src, phorbol esters, and growth factors. The present identification and purification of the S6 kinase should facilitate future studies aimed at elucidating the molecular mechanisms by which signals from these diverse stimuli rapidly converge upon and activate this enzyme.
Blenis
J
J
Department of Cellular and Developmental Biology, Harvard University, Cambridge, Massachusetts 02138.
Kuo
C J
CJ
Erikson
R L
RL
D000818:Animals; D002642:Chick Embryo; D006133:Growth Substances / Q000494:pharmacology*; D008970:Molecular Weight; D010703:Phorbol Esters / Q000494:pharmacology; D010766:Phosphorylation; D010770:Phosphotransferases / Q000032:analysis; D011494:Protein Kinases / Q000302:isolation & purification*; D017346:Protein Serine-Threonine Kinases; D038601:Ribosomal Protein S6; D012269:Ribosomal Proteins / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism
D006133:Growth Substances; D010703:Phorbol Esters; D038601:Ribosomal Protein S6; D012269:Ribosomal Proteins; D014443:Tyrosine; D010770:Phosphotransferases; D011494:Protein Kinases; D017346:Protein Serine-Threonine Kinases