Mitogen-responsive S6 kinase.
Many cell lines respond to mitogenic stimuli (serum, growth factors) with rapid phosphorylation of the ribosomal protein S6 at several serine sites. We have tried to identify the protein kinase(s) mediating this effect of growth stimuli. Examining post-DEAE chromatography fractions of S49 kin- cell extracts, we could detect a highly active effector-independent S6 kinase with specificity for serine residues. The study was extended to the presumably homologous human enzyme, using HeLa S3 cells as model system. Activity yields increased up to sevenfold when exhausted HeLa cells were supplied with fresh medium plus serum. The enzyme uses ATP, not GTP, as cosubstrate, 40-S or 80-S (reassociated from subunits) ribosomal particles being substrate. The optimal K+ concentration, measured at 3 mM Mg2+, is 35 mM. Under optimized assay conditions S6 phosphorylation proceeded faster in vitro than it appeared to do in vivo. The apparent Mr of the enzyme, as estimated by gel filtration on Sephadex G-100, is 56,000 (determination in the presence of 200 mM KCl in 25 mM phosphate buffer). Tighter binding to DEAE-Sephacel and higher specificity for S6 distinguishes this enzyme from the following S6-phosphorylating protein kinases: protein kinase C, protease-activated kinase II, histone-4 phosphotransferase and an enzyme with the properties of casein kinase I. In published summaries of observations shown here and in a follow-up study with chick embryo fibroblasts, the enzyme(s) has been referred to as mitogen-responsive S6 kinase(s) [Martini, O. H. W. and Lawen, A. (1985) in Hormones and cell regulation (Dumont, J. E., Hamprecht, B. and Nunez, J., eds) vol. 9, pp. 411-412, Elsevier Company, North-Holland, Amsterdam; Lawen, A. and Martini, O. H. W. (1985) FEBS Lett. 185, 272-276].
Lawen
A
A
Institut für Virologie und Immunbiologie, Universität Würzburg.
Burger
M
M
Martini
O H
OH
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D047388:Casein Kinases; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D008223:Lymphoma; D008934:Mitogens / Q000494:pharmacology*; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000494:pharmacology; D010766:Phosphorylation; D011188:Potassium / Q000494:pharmacology; D006656:Protamine Kinase / Q000378:metabolism; D011493:Protein Kinase C; D011494:Protein Kinases / Q000302:isolation & purification / Q000378:metabolism*; D038601:Ribosomal Protein S6; D019893:Ribosomal Protein S6 Kinases; D012269:Ribosomal Proteins / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D008934:Mitogens; D038601:Ribosomal Protein S6; D012269:Ribosomal Proteins; D000255:Adenosine Triphosphate; D000242:Cyclic AMP; D011494:Protein Kinases; D047388:Casein Kinases; D006656:Protamine Kinase; D019893:Ribosomal Protein S6 Kinases; D011493:Protein Kinase C; D010447:Peptide Hydrolases; D011188:Potassium