Regulation of protein kinase Cnu by the B-cell antigen receptor.
Diacylglycerol-dependent signaling plays an important role in signal transduction through T- and B-lymphocyte antigen receptors. Recently, a novel serine-threonine kinase of the protein kinase C (PKC) family has been described and designated as PKCnu. PKCnu has two putative diacylglycerol binding C1 domains, suggesting that it may participate in a novel diacylglycerol-mediated signaling pathway. Here we show that both endogenous and recombinant PKCnu are trans-located to the plasma membrane and activated by the diacylglycerol mimic phorbol 12-myristate 13-acetate. Mutational analysis demonstrates that PKCnu activation is dependent on trans-phosphorylation of two conserved activation loop serine residues. We also find that PKCnu is an important physiologic target of the B-cell receptor (BCR), because PKCnu is found to be abundantly expressed in chicken and human B-cell lines and, in addition, exhibits robust activation after BCR engagement. Genetic and pharmacologic analyses of BCR-mediated PKCnu activation indicate that it requires intact phospholipase Cgamma and PKC signaling pathways. Furthermore, in co-transfection assays, PKCnu can be trans-phosphorylated by the novel PKC isozymes PKCepsilon, PKCeta, or PKCtheta but not the classical PKC enzyme, PKCalpha. Taken together, these results suggest that PKCnu is an important component of signaling pathways downstream from novel PKC enzymes after B-cell receptor engagement.
Matthews
Sharon A
SA
Department of Pediatrics and Immunology, University of Washington and Children's Hospital and Regional Medical Center, Seattle, Washington 98195, USA.
Dayalu
Rashmi
R
Thompson
Lucas J
LJ
Scharenberg
Andrew M
AM
D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism; D015153:Blotting, Western; D020134:Catalytic Domain; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D006801:Humans; D051379:Mice; D008856:Microscopy, Fluorescence; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D011233:Precipitin Tests; D020033:Protein Isoforms; D011493:Protein Kinase C / Q000737:chemistry / Q000378:metabolism* / Q000502:physiology*; D011947:Receptors, Antigen, B-Cell / Q000378:metabolism*; D015398:Signal Transduction; D013347:Subcellular Fractions / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000378:metabolism; D014162:Transfection
D018076:DNA, Complementary; D000939:Epitopes; D020033:Protein Isoforms; D011947:Receptors, Antigen, B-Cell; C119190:protein kinase C nu; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate