Novel protein kinase C delta isoform insensitive to caspase-3.
Protein kinase C delta (PKC delta) plays a key regulatory role in a variety of cellular functions, including apoptosis, as well as cell growth and differentiation. We previously reported that apoptosis was induced by pretreatment with 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7), an inhibitor of PKC, in mouse thymocytes. In the present study, we showed that a novel PKC delta isoform (PKC deltaII) was transiently expressed when thymocytes were pretreated with H-7. The analysis of the cDNA encoding PKC deltaII indicated that a 78 bp fragment was inserted into the caspase-3 sensitive site of the original PKC delta (PKC deltaI), presumably by alternative splicing. The PKC deltaII expressed in COS-1 cells was one product with a molecular mass of 81 kDa and with kinase activity similar to that of PKC deltaI. The expressed PKC deltaI protein (78 kDa) was in part cleaved into a 38 kDa fragment in vivo and in vitro, but the PKC deltaII protein was not. Cleavage of the PKC deltaI protein was inhibited by a specific inhibitor of caspase-3, indicating that PKC deltaII is insensitive to caspase-3. The PKC deltaII was highly expressed in the testis and ovary, and at a lower level in the thymocytes, brain and kidney, whereas PKC deltaI was detected in most tissues, suggesting that the function of PKC deltaII is different from that of PKC deltaI.
Sakurai
Y
Y
Department of Biochemistry, Tokyo Dental College, Chiba, Japan.
Onishi
Y
Y
Tanimoto
Y
Y
Kizaki
H
H
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D019556:COS Cells; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism*; D002478:Cells, Cultured; D004251:DNA Transposable Elements; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D051745:Protein Kinase C-delta; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000187:drug effects / Q000201:enzymology
D004251:DNA Transposable Elements; D004791:Enzyme Inhibitors; D007527:Isoenzymes; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; C497259:Prkcd protein, mouse; D011493:Protein Kinase C; D051745:Protein Kinase C-delta; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases