title: Protein kinase C delta (PKCdelta) inhibits the expression of glutamine synthetase in glial cells via the PKCdelta regulatory domain and its tyrosine phosphorylation.
abstract: Protein kinase C (PKC) plays an important role in the proliferation and differentiation of glial cells. In a recent study we found that overexpression of PKCdelta reduced the expression of the astrocytic marker glutamine synthetase (GS). In this study we explored the mechanisms involved in the inhibitory effect of PKCdelta on the expression of glutamine synthetase. Using PKC chimeras we first examined the role of the catalytic and regulatory domains of PKCdelta on the expression of glutamine synthetase. We found that cells stably transfected with chimeras between the regulatory domain of PKCdelta and the catalytic domains of PKCalpha or epsilon inhibited the expression of GS, similar to the inhibition exerted by overexpression of PKCdelta itself. In contrast, no significant effects were observed in cells transfected with the reciprocal PKC chimeras between the regulatory domains of PKCalpha or epsilon and the catalytic domain of PKCdelta. PKCdelta has been shown to undergo tyrosine phosphorylation in response to various activators. Tyrosine phosphorylation of PKCdelta in response to phorbol 12-myristate 13-acetate and platelet-derived growth factor occurred only in chimeras which contained the PKCdelta regulatory domain. Cells transfected with a PKCdelta mutant (PKCdelta5), in which the five putative tyrosine phosphorylation sites were mutated to phenylalanine, showed markedly diminished tyrosine phosphorylation in response to phorbol 12-myristate 13-acetate and platelet-derived growth factor and normal levels of GS. Our results indicate that the regulatory domain of PKCdelta mediates the inhibitory effect of this isoform on the expression of GS. Phosphorylation of PKCdelta on tyrosine residues in the regulatory domain is implicated in this inhibitory effect.
authors.affiliation: Molecular Mechanisms of Tumor Promotion Section, Laboratory of Cellular Carcinogenesis and Tumor Promotion, NCI, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: C
authors.initials: C
authors.lastname: Brodie
authors.forename: K
authors.initials: K
authors.lastname: Bogi
authors.forename: P
authors.initials: P
authors.lastname: Acs
authors.forename: P S
authors.initials: PS
authors.lastname: Lorenzo
authors.forename: L
authors.initials: L
authors.lastname: Baskin
authors.forename: P M
authors.initials: PM
authors.lastname: Blumberg
mesh_terms: D054713:Bryostatins; D020134:Catalytic Domain / Q000235:genetics; D002478:Cells, Cultured; D004789:Enzyme Activation; D015971:Gene Expression Regulation, Enzymologic; D005974:Glutamate-Ammonia Ligase / Q000235:genetics*; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D007783:Lactones / Q000494:pharmacology; D018942:Macrolides; D016297:Mutagenesis, Site-Directed; D009457:Neuroglia / Q000201:enzymology*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism*; D051745:Protein Kinase C-delta; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014443:Tyrosine / Q000378:metabolism*
chemical_list: D054713:Bryostatins; D007527:Isoenzymes; D007783:Lactones; D018942:Macrolides; D011993:Recombinant Fusion Proteins; C046785:bryostatin 1; D014443:Tyrosine; C497258:PRKCD protein, human; D011493:Protein Kinase C; D051745:Protein Kinase C-delta; D005974:Glutamate-Ammonia Ligase; D013755:Tetradecanoylphorbol Acetate