Phosphatidylserine externalization during CD95-induced apoptosis of cells and cytoplasts requires ICE/CED-3 protease activity.
Phosphatidylserine (PS), a lipid normally confined to the inner leaflet of the plasma membrane, is exported to the outer plasma membrane leaflet during apoptosis to serve as a trigger for recognition of apoptotic cells by phagocytes. The mechanism of PS export during apoptosis is not known nor is it clear whether the nuclear changes that typify apoptosis contribute in any way to this event. Here, we demonstrate that ligation of the CD95 (Fas/APO-1) molecule on Jurkat cytoplasts induces dramatic PS externalization similar to that observed during apoptosis of intact cells. Apoptosis of both cells and cytoplasts was associated with proteolytic processing of CPP32, a member of the interleukin-1beta converting enzyme (ICE)/CED-3 protease family, to its active form. Fodrin, a component of the cortical cytoskeleton, also underwent proteolytic cleavage during apoptosis of both cytoplasts and intact cells. Strikingly, CPP32 activation, fodrin proteolysis, and PS externalization were all inhibited in the presence of peptide inhibitors of ICE/CED-3 family proteases. These data provide strong support for the notion that the cell death machinery is extranuclear and is likely to be comprised of one or more members of the ICE/CED-3 family and that activation of this machinery does not require nuclear participation.
Martin
S J
SJ
Division of Cellular Immunology, La Jolla Institute for Allergy and Immunology, San Diego, California 92121, USA. sjmartin@may.ie
Finucane
D M
DM
Amarante-Mendes
G P
GP
O'Brien
G A
GA
Green
D R
DR
D017209:Apoptosis / Q000187:drug effects; D029742:Caenorhabditis elegans Proteins; D020170:Caspase 1; D020169:Caspases; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D015801:Helminth Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006801:Humans; D019169:Jurkat Cells; D010718:Phosphatidylserines / Q000378:metabolism*; D019014:fas Receptor / Q000378:metabolism*
D029742:Caenorhabditis elegans Proteins; D015853:Cysteine Proteinase Inhibitors; D015801:Helminth Proteins; D010718:Phosphatidylserines; D019014:fas Receptor; D020169:Caspases; D003546:Cysteine Endopeptidases; C089223:ced-3 protein, C elegans; D020170:Caspase 1