Pancreastatin, a chromogranin A-derived peptide, activates protein synthesis signaling cascade in rat adipocytes.
Pancreastatin (PST), a chromogranin A-derived peptide, has been found to modulate glucose, lipid, and protein metabolism in rat adipocytes. PST has an overall counterregulatory effect on insulin action by activating a specific receptor-effector system (Galpha(q/11) protein-PLC-beta-PKC(classical)). However, PST stimulates both basal and insulin-mediated protein synthesis in rat adipocytes. In order to further investigate the mechanisms underlying the effect of PST stimulating protein synthesis, we sought to study the regulation of different components of the core translational machinery by the signaling triggered by PST. Thus, we studied ribosomal p70 S6 kinase, phosphorylation of the cap-binding protein (initiation factor) eIF4E, and phosphorylation of the eIF4E-binding protein 4E-BP1 (PHAS-I). We have found that PST stimulates the S6 kinase activity, as assessed by kinase assay using specific immunoprecipitates and substrate. This effect was checked by Western blot with specific antibodies against the phosphorylated S6 kinase. Thus, PST dose-dependently stimulates Thr421/Ser424 phosphorylation of S6 kinase. Moreover, PST promotes phosphorylation of regulatory sites in 4E-BP1 (PHAS-I) (Thr37, Thr46). The initiation factor eIF4E itself, whose activity is also increased upon phosphorylation, is phosphorylated in Ser209 by PST stimulation. Finally, we have found that these effects of PST on S6 kinase and the translation machinery can be blocked by preventing the activation of PKC. These results indicate that PST stimulates protein synthesis machinery by activating PKC and provides some evidence of the molecular mechanisms involved, i.e., the activation of S6K and the phosphorylation of 4E-BP1 (PHAS-I) and the initiation factor eIF4E.
González-Yanes
Carmen
C
Department of Medical Biochemistry and Molecular Biology, Medical School, Investigation Unit, Virgen Macarena University Hospital, Av. Sánchez Pizjuan, 4. Seville 41009, Spain.
Sánchez-Margalet
Víctor
V
D017667:Adipocytes / Q000166:cytology / Q000502:physiology*; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism; D053379:Chromogranin A; D002864:Chromogranins / Q000378:metabolism; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000378:metabolism; D039561:Eukaryotic Initiation Factor-4E / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D008297:Male; D010187:Pancreatic Hormones / Q000378:metabolism*; D010448:Peptide Initiation Factors / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D014176:Protein Biosynthesis; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D038762:Ribosomal Protein S6 Kinases, 70-kDa / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*
D002352:Carrier Proteins; D053379:Chromogranin A; D002864:Chromogranins; C499526:Eif4ebp1 protein, rat; D004791:Enzyme Inhibitors; D039561:Eukaryotic Initiation Factor-4E; D047908:Intracellular Signaling Peptides and Proteins; D010187:Pancreatic Hormones; D010448:Peptide Initiation Factors; D010750:Phosphoproteins; C506153:chromogranin A, rat; C050836:pancreastatin; D038762:Ribosomal Protein S6 Kinases, 70-kDa; D011493:Protein Kinase C