Characterization of a serine protease-mediated cell death program activated in human leukemia cells.
Tightly controlled proteolysis is a defining feature of apoptosis and caspases are critical in this regard. Significant roles for non-caspase proteases in cell death have been highlighted. Staurosporine causes a rapid induction of apoptosis in virtually all mammalian cell types. Numerous studies demonstrate that staurosporine can activate cell death under caspase-inhibiting circumstances. The aim of this study was to investigate the proteolytic mechanisms responsible for cell death under these conditions. To that end, we show that inhibitors of serine proteases can delay cell death in one such system. Furthermore, through profiling of proteolytic activation, we demonstrate, for the first time, that staurosporine activates a chymotrypsin-like serine protease-dependent cell death in HL-60 cells independently, but in parallel with the caspase controlled systems. Features of the serine protease-mediated system include cell shrinkage and apoptotic morphology, regulation of caspase-3, altered nuclear morphology, generation of an endonuclease and DNA degradation. We also demonstrate a staurosporine-induced activation of a putative 16 kDa chymotrypsin-like protein during apoptosis.
O'Connell
A R
AR
National Centre for Biomedical Engineering Science and Department of Biochemistry, National University of Ireland, Galway, Ireland.
Holohan
C
C
Torriglia
A
A
Lee
B W
BW
Lee
B F
BF
Stenson-Cox
C
C
D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D053148:Caspase 3; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism; D053818:Chymases; D018922:HL-60 Cells / Q000201:enzymology / Q000473:pathology; D006801:Humans; D012697:Serine Endopeptidases / Q000378:metabolism / Q000502:physiology*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology*; D019311:Staurosporine / Q000494:pharmacology; D013347:Subcellular Fractions
D061945:Caspase Inhibitors; D015842:Serine Proteinase Inhibitors; D012697:Serine Endopeptidases; D053818:Chymases; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D019311:Staurosporine