Modulation of zeta-protein kinase C by cyclic AMP in PC12 cells occurs through phosphorylation by protein kinase A.
Although cyclic AMP (cAMP) has been reported to cross talk with the protein kinase C (PKC) system, effects of elevated intracellular cAMP on the activities of specific PKC isoforms have not been studied. We report findings from a permeabilized cell assay that was used to examine changes in the activity of the atypical PKC isoforms brought about by exposure of PC12 cells to agents that elevate intracellular cAMP. We found that increases in intracellular cAMP led to rapid stimulation of atypical PKC activity, 40-70% above control, for a sustained period of time, a response that occurred independent of the phorbol 12-myristate 13-acetate (PMA)-sensitive PKC isoforms. Changes in intracellular cAMP levels resulted in a dose-dependent redistribution of zeta-PKC to the cytoplasm with a concomitant increase in the phosphorylation state of the enzyme. Incubation of purified zeta-PKC with increasing concentrations of PKA likewise caused a twofold increase in the phosphorylation state of zeta-PKC. In contrast to the positive effect that PKA-mediated phosphorylation had on the activity of zeta-PKC, the enzyme displayed reduced binding to ras when phosphorylated. Taken together, these findings are consistent with the hypothesis that protein phosphorylation of PKC acts as a positive effector of its enzyme activity and may serve as a negative modulator for interaction with other proteins.
Wooten
M W
MW
Department of Zoology, Auburn University, AL 36849-5414, USA.
Seibenhener
M L
ML
Matthews
L H
LH
Zhou
G
G
Coleman
E S
ES
D000818:Animals; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004789:Enzyme Activation / Q000502:physiology; D007527:Isoenzymes / Q000378:metabolism*; D016716:PC12 Cells / Q000201:enzymology*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats; D018411:Spodoptera / Q000166:cytology; D018631:ras Proteins / Q000378:metabolism; D000099221:Protein Kinase C zeta
D017871:Calcium-Calmodulin-Dependent Protein Kinases; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D007527:Isoenzymes; D011493:Protein Kinase C; D018631:ras Proteins; D000099221:Protein Kinase C zeta