Calcineurin is required for translational control of protein synthesis in rat pancreatic acini.
CCK increases the rate of net protein synthesis in rat pancreatic acini by activating initiation and elongation factors required for translation. The immunosuppressant FK506 inhibits the Ca(2+)-calmodulin-dependent phosphatase calcineurin in pancreatic acinar cells and blocks pancreatic growth induced by chronic CCK treatment. To test a requirement for calcineurin in the activation of the translational machinery stimulated by CCK, we evaluated the effects of FK506 on protein synthesis and on regulatory initiation and elongation factors in rat pancreatic acini in vitro. CCK acutely increased protein synthesis in acini from normal rats with a maximum increase at 100 pM CCK to 170 +/- 11% of control. The immunosuppressant FK506 dose-dependently inhibited CCK-stimulated protein synthesis over the same concentration range that blocked calcineurin activity, as assessed by dephosphorylation of the calcineurin substrate calcium-regulated heat-stable protein of 24 kDa. Another immunosuppressant, cyclosporin A, inhibited protein synthesis, but its effects appeared more complex. FK506 also inhibited protein synthesis stimulated by bombesin and carbachol. FK506 did not significantly affect the activity of the initiation factor-2B, or the phosphorylation of the initiation factor-2alpha, ribosomal protein protein S6, or the mRNA cap binding protein eukaryotic initiation factor (eIF) 4E. Instead, blockade of calcineurin with FK506 reduced the phosphorylation of the eIF4E binding protein, reduced the formation of the eIF4F complex, and increased the phosphorylation of eukaryotic elongation factor 2. From these results, we conclude that calcineurin activity is required for protein synthesis, and this action may be related to an effect on the formation of the mRNA cap binding complex and the elongation processes.
Sans
Maria Dolors
MD
Department of Molecular and Integrative Physiology, University of Michigan Medical School, 1301 E. Catherine St., 7737 Med Sci II, Ann Arbor, MI 48109-0622, USA. mdsansg@umich.edu
Williams
John A
JA
D000818:Animals; D019703:Calcineurin / Q000378:metabolism*; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism; D002766:Cholecystokinin / Q000494:pharmacology; D016572:Cyclosporine / Q000494:pharmacology; D054736:Elongation Factor 2 Kinase; D020717:Eukaryotic Initiation Factor-2B / Q000378:metabolism; D039561:Eukaryotic Initiation Factor-4E; D039562:Eukaryotic Initiation Factor-4F / Q000378:metabolism; D007166:Immunosuppressive Agents / Q000494:pharmacology; D047908:Intracellular Signaling Peptides and Proteins; D008297:Male; D010179:Pancreas / Q000502:physiology*; D020652:Peptide Elongation Factor 2 / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D014176:Protein Biosynthesis / Q000187:drug effects / Q000502:physiology*; D039381:RNA Cap-Binding Proteins / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D038601:Ribosomal Protein S6 / Q000378:metabolism; D016559:Tacrolimus / Q000494:pharmacology
D002352:Carrier Proteins; C112609:EIF4E2 protein, human; C499526:Eif4ebp1 protein, rat; D020717:Eukaryotic Initiation Factor-2B; D039561:Eukaryotic Initiation Factor-4E; D039562:Eukaryotic Initiation Factor-4F; D007166:Immunosuppressive Agents; D047908:Intracellular Signaling Peptides and Proteins; D020652:Peptide Elongation Factor 2; D010750:Phosphoproteins; D039381:RNA Cap-Binding Proteins; D038601:Ribosomal Protein S6; D016572:Cyclosporine; D002766:Cholecystokinin; C517689:EEF2K protein, human; C517691:Eef2k protein, rat; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D054736:Elongation Factor 2 Kinase; D019703:Calcineurin; D016559:Tacrolimus