Caspase-mediated inhibition of human cytosolic phospholipase A2 during apoptosis.
Activation of cytosolic phospholipase A2 (cPLA2) is an essential step in the initiation of the cascade of enzymatic reactions leading to the generation of proinflammatory lipid mediators. Hence, the regulation of cPLA2 is a key event in the induction of inflammatory responses. cPLA2 is activated, in part, by apoptotic stimuli such as TNF or Fas ligand. Apoptosis, however, does not provoke an inflammatory response. Here, we demonstrate that cPLA2 is cleaved by caspase-3 and/or a related caspase in HeLa cells undergoing apoptosis. Mutation of a predicted caspase-3 cleavage site abolishes cPLA2 processing both in vitro and in intact cells. The 70-kDa cleavage product of cPLA2 itself has no catalytic function, while inhibition of cleavage results in an increased enzymatic activity. Additionally, overexpression of the 70-kDa fragment appears to produce a dominant negative effect on endogenous cPLA2 activity. In HeLa cells, cPLA2 activity was dispensable for the course of apoptosis. We cannot rule out, however, that cPLA2 activity is involved in the induction of apoptosis in other cell types. Taken together, our results suggest that the enzymatic activity of cPLA2 is specifically inhibited by caspase-mediated cleavage during apoptosis. The inactivation of cPLA2 represents a previously unrecognized mechanism for avoiding an inflammatory reaction against apoptotic cells.
Adam-Klages
S
S
Institut für Immunologie, Christian-Albrechts-Universität Kiel, Germany. sabine@immunologie.uni-kiel.de
Schwandner
R
R
Lüschen
S
S
Ussat
S
S
Kreder
D
D
Krönke
M
M
D017209:Apoptosis / Q000235:genetics; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism / Q000502:physiology*; D002384:Catalysis; D002460:Cell Line; D003600:Cytosol / Q000201:enzymology*; D004252:DNA Mutational Analysis; D006367:HeLa Cells; D006801:Humans; D006868:Hydrolysis; D007249:Inflammation / Q000201:enzymology / Q000473:pathology; D007668:Kidney / Q000166:cytology; D016297:Mutagenesis, Site-Directed; D010741:Phospholipases A / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D054467:Phospholipases A2
D010741:Phospholipases A; D054467:Phospholipases A2; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases