Role of protein kinase C and NF-kappaB in proteolysis-inducing factor-induced proteasome expression in C(2)C(12) myotubes.
Proteolysis-inducing factor (PIF) is a sulphated glycoprotein produced by cachexia-inducing tumours, which initiates muscle protein degradation through an increased expression of the ubiquitin-proteasome proteolytic pathway. The role of kinase C (PKC) in PIF-induced proteasome expression has been studied in murine myotubes as a surrogate model of skeletal muscle. Proteasome expression induced by PIF was attenuated by 4alpha-phorbol 12-myristate 13-acetate (100 nM) and by the PKC inhibitors Ro31-8220 (10 microM), staurosporine (300 nM), calphostin C (300 nM) and Gö 6976 (200 microM). Proteolysis-inducing factor-induced activation of PKC(alpha), with translocation from the cytosol to the membrane at the same concentration as that inducing proteasome expression, and this effect was attenuated by calphostin C. Myotubes transfected with a constitutively active PKC(alpha) (pCO(2)) showed increased expression of proteasome activity, and a longer time course, compared with their wild-type counterparts. In contrast, myotubes transfected with a dominant-negative PKC(alpha) (pKS1), which showed no activation of PKC(alpha) in response to PIF, exhibited no increase in proteasome activity at any time point. Proteolysis-inducing factor-induced proteasome expression has been suggested to involve the transcription factor nuclear factor-kappaB (NF-kappaB), which may be activated through PKC. Proteolysis-inducing factor induced a decrease in cytosolic I-kappaBalpha and an increase in nuclear binding of NF-kappaB in pCO(2), but not in pKS1, and the effect in wild-type cells was attenuated by calphostin C, confirming that it was mediated through PKC. This suggests that PKC may be involved in the phosphorylation and degradation of I-kappaBalpha, induced by PIF, necessary for the release of NF-kappaB from its inactive cytosolic complex.
Smith
H J
HJ
Pharmaceutical Sciences Research Institute, Aston University, Birmingham, B4 7ET, UK.
Wyke
S M
SM
Tisdale
M J
MJ
D000818:Animals; D001798:Blood Proteins / Q000494:pharmacology*; D015153:Blotting, Western; D002478:Cells, Cultured; D003546:Cysteine Endopeptidases / Q000096:biosynthesis* / Q000187:drug effects; D024202:Electrophoretic Mobility Shift Assay; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D020672:I-kappa B Proteins / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D009097:Multienzyme Complexes / Q000096:biosynthesis* / Q000187:drug effects; D018485:Muscle Fibers, Skeletal / Q000378:metabolism*; D016328:NF-kappa B / Q000187:drug effects / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex; D011493:Protein Kinase C / Q000187:drug effects / Q000378:metabolism*; D021381:Protein Transport; D011509:Proteoglycans; D014162:Transfection
D001798:Blood Proteins; D004791:Enzyme Inhibitors; D020672:I-kappa B Proteins; D009097:Multienzyme Complexes; D016328:NF-kappa B; D011509:Proteoglycans; C042057:proteolysis-inducing peptide; D011493:Protein Kinase C; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex