Signal transduction pathways involved in proteolysis-inducing factor induced proteasome expression in murine myotubes.
The proteolysis-inducing factor (PIF) is produced by cachexia-inducing tumours and initiates protein catabolism in skeletal muscle. The potential signalling pathways linking the release of arachidonic acid (AA) from membrane phospholipids with increased expression of the ubiquitin-proteasome proteolytic pathway by PIF has been studied using C(2)C(12) murine myotubes as a surrogate model of skeletal muscle. The induction of proteasome activity and protein degradation by PIF was blocked by quinacrine, a nonspecific phospholipase A(2) (PLA(2)) inhibitor and trifluroacetyl AA, an inhibitor of cytosolic PLA(2). PIF was shown to increase the expression of calcium-independent cytosolic PLA(2), determined by Western blotting, at the same concentrations as those inducing maximal expression of 20S proteasome alpha-subunits and protein degradation. In addition, both U-73122, which inhibits agonist-induced phospholipase C (PLC) activation and D609, a specific inhibitor of phosphatidylcholine-specific PLC also inhibited PIF-induced proteasome activity. This suggests that both PLA(2) and PLC are involved in the release of AA in response to PIF, and that this is important in the induction of proteasome expression. The two tyrosine kinase inhibitors genistein and tryphostin A23 also attenuated PIF-induced proteasome expression, implicating tyrosine kinase in this process. PIF induced phosphorylation of p44/42 mitogen-activated protein kinase (MAPK) at the same concentrations as that inducing proteasome expression, and the effect was blocked by PD98059, an inhibitor of MAPK kinase, as was also the induction of proteasome expression, suggesting a role for MAPK activation in PIF-induced proteasome expression.
Pharmaceutical Sciences Research Institute, Aston University, Birmingham B4 7ET, UK.
H J
HJ
Smith
M J
MJ
Tisdale
D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D001798:Blood Proteins / Q000494:pharmacology*; D015153:Blotting, Western; D003546:Cysteine Endopeptidases / Q000096:biosynthesis / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000187:drug effects / Q000378:metabolism; D009097:Multienzyme Complexes / Q000096:biosynthesis / Q000187:drug effects*; D018485:Muscle Fibers, Skeletal / Q000187:drug effects* / Q000378:metabolism; D018482:Muscle, Skeletal / Q000502:physiology; D010741:Phospholipases A / Q000187:drug effects / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex; D011509:Proteoglycans; D015398:Signal Transduction / Q000502:physiology*; D010738:Type C Phospholipases / Q000187:drug effects / Q000378:metabolism
D001798:Blood Proteins; D004791:Enzyme Inhibitors; D009097:Multienzyme Complexes; D011509:Proteoglycans; C042057:proteolysis-inducing peptide; D016718:Arachidonic Acid; D020928:Mitogen-Activated Protein Kinases; D010741:Phospholipases A; D010738:Type C Phospholipases; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex