VP-16 (etoposide) and calphostin C trigger different nuclear but akin cytoplasmic patterns of changes in the distribution and activity of protein kinase C-betaI in polyomavirus-transformed pyF111 rat fibroblasts.
Protein kinase C (PKC) isoforms regulate cell proliferation and apoptosis. Since the PKC isoenzyme complement varies considerably from cell type to cell type, a PKC's responsiveness to an apoptogenic challenge must be defined for both the type of apoptogen and the type of cell. We have already reported that the changes in the distribution and activity of PKC-delta in apoptosing polyomavirus-infected/transformed Fischer rat embryo pyF111 fibroblasts depend on the type of apoptogen. Here, we show that this is also true for PKC-betaI in pyF111 cells treated with the slow DNA-damaging VP-16 (etoposide) or the fast-acting (in the cytoplasm) calphostin C. These apoptogens caused quite different shifts of the PKC-betaI level and activity in the nuclear membrane (NM) and nucleoplasm (NP), but corresponding changes in the cytosol (CS) and cytoplasmic particulate (CP) fractions. The hefty translocation of PKC-betaI onto the CP fraction and its increased activity there suggest the possible triggering of a cytochrome c/caspase-mediated apoptosis-inducing mechanism common to both agents. The present results are a necessary lead-up to functional proteomic analyses aimed at identifying the molecules forming the local PKC-betaI signalling modules under different conditions.
Chiarini
Anna
A
Histology and Embryology Unit, Department of Biomedical and Surgical Sciences, University of Verona Medical School, Verona I-37134, Italy.
Whitfield
James F
JF
Armato
Ubaldo
U
Dal Pra
Ilaria
I
D000818:Animals; D017209:Apoptosis / Q000502:physiology; D002460:Cell Line; D002467:Cell Nucleus / Q000201:enzymology; D002472:Cell Transformation, Viral; D003593:Cytoplasm / Q000201:enzymology; D004791:Enzyme Inhibitors / Q000378:metabolism; D005047:Etoposide / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D007527:Isoenzymes / Q000378:metabolism; D009281:Naphthalenes / Q000378:metabolism*; D019384:Nucleic Acid Synthesis Inhibitors / Q000378:metabolism; D011120:Polyomavirus / Q000235:genetics / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism*; D064546:Protein Kinase C beta; D051381:Rats; D011916:Rats, Inbred F344
D004791:Enzyme Inhibitors; D007527:Isoenzymes; D009281:Naphthalenes; D019384:Nucleic Acid Synthesis Inhibitors; D005047:Etoposide; D011493:Protein Kinase C; D064546:Protein Kinase C beta; C058819:calphostin C