title: Fas-induced activation of the cell death-related protease CPP32 Is inhibited by Bcl-2 and by ICE family protease inhibitors.
abstract: The human proto-oncogene bcl-2 and its Caenorhabditis elegans homologue ced-9 inhibit programmed cell death. In contrast, members of the human interleukin-1beta converting enzyme (ICE) family of cysteine proteases and their C. elegans homologue CED-3 promote the death program. Genetic experiments in C. elegans have shown that ced-9 is formally a negative regulator of ced-3 function, but neither those studies nor others have determined whether CED-9 or Bcl-2 proteins act biochemically upstream or downstream of CED-3/ICE proteases. CPP32, like all known members of the CED-3/ICE family, is synthesized as a proenzyme that is subsequently processed into an active protease with specificity for cleavage at Asp-X peptide bonds. In this report, we demonstrate that the CPP32 proenzyme is proteolytically processed and activated in Jurkat cells induced to die by Fas ligation. CPP32 activation is blocked by cell-permeable inhibitors of aspartate-directed, cysteine proteases, suggesting that pro-CPP32 is cleaved by active CPP32 or by other ICE family members. Heterologous expression of Bcl-2 in Jurkat cells prevents Fas-induced cell death as well as proteolytic processing and activation of CPP32. Thus, Bcl-2 acts at or upstream of the CPP32 activation step to inhibit apoptosis induced by Fas stimulation.
authors.affiliation: IDUN Pharmaceuticals, Inc., La Jolla, California 92037, USA.
authors.forename: R C
authors.initials: RC
authors.lastname: Armstrong
authors.forename: T
authors.initials: T
authors.lastname: Aja
authors.forename: J
authors.initials: J
authors.lastname: Xiang
authors.forename: S
authors.initials: S
authors.lastname: Gaur
authors.forename: J F
authors.initials: JF
authors.lastname: Krebs
authors.forename: K
authors.initials: K
authors.lastname: Hoang
authors.forename: X
authors.initials: X
authors.lastname: Bai
authors.forename: S J
authors.initials: SJ
authors.lastname: Korsmeyer
authors.forename: D S
authors.initials: DS
authors.lastname: Karanewsky
authors.forename: L C
authors.initials: LC
authors.lastname: Fritz
authors.forename: K J
authors.initials: KJ
authors.lastname: Tomaselli
mesh_terms: D000595:Amino Acid Sequence; D017209:Apoptosis; D020170:Caspase 1; D053148:Caspase 3; D020169:Caspases; D002460:Cell Line; D003546:Cysteine Endopeptidases / Q000187:drug effects* / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology*; D004789:Enzyme Activation; D006801:Humans; D006868:Hydrolysis; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D019253:Proto-Oncogene Proteins c-bcl-2; D019014:fas Receptor / Q000378:metabolism*
chemical_list: D015853:Cysteine Proteinase Inhibitors; C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D019014:fas Receptor; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D003546:Cysteine Endopeptidases; D020170:Caspase 1