Modulation of PKCδ tyrosine phosphorylation and activity in salivary and PC-12 cells by Src kinases

Abstract

Protein kinase C (PKC) δ becomes tyrosine phosphorylated in rat parotid acinar cells exposed to muscarinic and substance P receptor agonists, which initiate fluid secretion in this salivary cell. Here we examine the signaling components of PKCδ tyrosine phosphorylation and effects of phosphorylation on PKCδ activity. Carbachol- and substance P-promoted increases in PKCδ tyrosine phosphorylation were blocked by inhibiting phospholipase C (PLC) but not by blocking intracellular Ca2+ concentration elevation, suggesting that diacylglycerol, rather than D-myo-inositol 1,4,5-trisphosphate production, positively modulated this phosphorylation. Stimuli-dependent increases in PKCδ activity in parotid and PC-12 cells were blocked in vivo by inhibitors of Src tyrosine kinases. Dephosphorylation of tyrosine residues by PTP1B, a protein tyrosine phosphatase, reduced the enhanced PKCδ activity. Lipid cofactors modified the tyrosine phosphorylation-dependent PKCδ activation. Two PKCδ regulatory sites (Thr-505 and Ser-662) were constitutively phosphorylated in unstimulated parotid cells, and these phosphorylations were not altered by stimuli that increased PKCδ tyrosine phosphorylation. These results demonstrate that PKCδ activity is positively modulated by tyrosine phosphorylation in parotid and PC-12 cells and suggest that PLC-dependent effects of secretagogues on salivary cells involve Src-related kinases.