The phosphorylation of tyrosine 332 is necessary for the caspase 3-dependent cleavage of PKCdelta and the regulation of cell apoptosis.
Protein kinase C delta (PKCdelta plays a major role in the regulation of cell apoptosis and survival. PKCdelta is cleaved by caspase 3 to generate a constitutively active catalytic domain that mediates both its apoptotic and anti-apoptotic effects. The caspase cleavage site of PKCdelta in the hinge region is flanked by the two tyrosine residues, Y311 and Y332. Here, we examined the role of the phosphorylation of tyrosines 311 and 332 in the cleavage and apoptotic function of PKCdelta using the apoptotic stimuli, TRAIL and cisplatin. Tyrosine 332 was constitutively phosphorylated in the A172 and HeLa cells and was further phosphorylated by TRAIL and cisplatin. This phosphorylation was inhibited by the Src inhibitors, PP2 and SU6656, and by silencing of Src. Treatment of the A172 and HeLa cells with TRAIL induced cleavage of the WT PKCdelta and of the PKCdeltaY311F mutant, whereas a lower level of cleavage was observed in the PKCdeltaY332F mutant. Similarly, a smaller degree of cleavage of the PKCdeltaY332 mutant was observed in LNZ308 cells treated with cisplatin. Mutation of Y332F affected the apoptotic function of PKCdelta; overexpression of the PKCdeltaY332 mutant increased the apoptotic effect of TRAIL, whereas it decreased the apoptotic effect of cisplatin. Inhibition of Src decreased the cleavage of PKCdelta and modified the apoptotic responses of the cells to TRAIL and cisplatin, similar to effect of the PKCdeltaY332F mutant. These results demonstrate that the phosphorylation of tyrosine 332 by Src modulates the cleavage of PKCdelta and the sensitivity of glioma cells to TRAIL and cisplatin.
William and Karen Davidson Laboratory of Cell Signaling and Tumorigenesis, Hermelin Brain Tumor Center, Department of Neurosurgery, Henry Ford Hospital, Detroit, MI, USA.
Wei
W
Lu
Hae-Kyung
HK
Lee
Cunli
C
Xiang
Susan
S
Finniss
Chaya
C
Brodie
D019943:Amino Acid Substitution; D017209:Apoptosis; D053148:Caspase 3 / Q000378:metabolism*; D002460:Cell Line; D002945:Cisplatin / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D010766:Phosphorylation; D051745:Protein Kinase C-delta / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000378:metabolism; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000494:pharmacology; D014443:Tyrosine / Q000378:metabolism*
D053221:TNF-Related Apoptosis-Inducing Ligand; D014443:Tyrosine; D016392:Proto-Oncogene Proteins pp60(c-src); D051745:Protein Kinase C-delta; D053148:Caspase 3; D002945:Cisplatin