p70s6k is activated by CCK in rat pancreatic acini.
The expression and activity of p70s6k-p85s6k in isolated rat pancreatic acini were revealed by Western blotting, immunoprecipitation, and kinase assay. Cholecystokinin (CCK) stimulation of p70s6k activity was biphasic, with an early phase maximum at 5 min and a late phase maximum at 60 min. The threshold concentration of CCK to increase p70s6k activity was 3 pM, and the maximal effect was seen at 1 nM CCK. Carbachol and bombesin, but not vasoactive intestinal peptide, also activated p70s6k. The protein kinase C (PKC) activator (12-O-tetradecanoylphorbol 13-acetate), the calcium ionophore (ionomycin), and a derivative of adenosine 3',5'-cyclic monophosphate induced only a slight increase in p70s6k activity. Rapamycin potently blocked both the basal and the CCK-stimulated p70s6k activity, and this inhibition was reversed by an excess of FK-506. The phosphatidylinositol 3-kinase inhibitor, wortmannin, potently inhibited p70s6k activation by CCK, whereas the tyrosine kinase inhibitor genistein had only a partial effect. Neither rapamycin nor wortmannin inhibited amylase release at concentrations that inhibited p70s6k activity. Thus the activation pathway of p70s6k by CCK is not mediated by PKC or mobilization of intracellular calcium but seems to be mediated by phosphatidylinositol 3-kinase. The effect of rapamycin to inhibit p70s6k activity is mediated by binding to the immunophyllin FK-506-binding protein of 12 kDa. The p70s6k is not involved in the secretion of digestive enzymes induced by CCK.
Bragado
M J
MJ
Department of Physiology, University of Michigan, Ann Arbor 48109-0622, USA.
Groblewski
G E
GE
Williams
J A
JA
D000595:Amino Acid Sequence; D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D001839:Bombesin / Q000494:pharmacology; D002217:Carbachol / Q000494:pharmacology; D002478:Cells, Cultured; D002766:Cholecystokinin / Q000494:pharmacology*; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000494:pharmacology; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019833:Genistein; D015759:Ionomycin / Q000494:pharmacology; D007529:Isoflavones / Q000494:pharmacology; D007700:Kinetics; D008297:Male; D008969:Molecular Sequence Data; D010179:Pancreas / Q000201:enzymology*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000037:antagonists & inhibitors; D011090:Polyenes / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D019893:Ribosomal Protein S6 Kinases; D020123:Sirolimus; D013379:Substrate Specificity; D016559:Tacrolimus / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013873:Thionucleotides / Q000494:pharmacology; D014660:Vasoactive Intestinal Peptide / Q000494:pharmacology; D000077191:Wortmannin
D000730:Androstadienes; D001839:Bombesin; D002217:Carbachol; D002766:Cholecystokinin; D000242:Cyclic AMP; D004791:Enzyme Inhibitors; D019833:Genistein; D015759:Ionomycin; D007529:Isoflavones; D010455:Peptides; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor); D011090:Polyenes; D017346:Protein Serine-Threonine Kinases; D019893:Ribosomal Protein S6 Kinases; D020123:Sirolimus; D016559:Tacrolimus; D013755:Tetradecanoylphorbol Acetate; D013873:Thionucleotides; D014660:Vasoactive Intestinal Peptide; D000077191:Wortmannin; C015929:8-((4-chlorophenyl)thio)cyclic-3',5'-AMP