Sequential and rapid activation of select caspases during apoptosis of normal intestinal epithelial cells.
Detachment-induced cell death (DICD) is considered to be one of the means by which intestinal epithelial cells (IEC) die of apoptosis as they reach the lumen and are shed. Caspases, a family of cysteine proteases, play a central role in initiating, amplifying, and executing apoptosis; however, the pattern of caspase activation in response to distinct apoptotic stimuli remains unknown. We investigated the kinetics of caspase activation during DICD in freshly isolated human IEC. DNA fragmentation is observed 90 min after detachment and is preceded by the sequential activation of preformed members of the CPP32 family of caspases. Activation of caspase 6 and cleavage of the endogenous caspase substrate poly(ADP-ribose) polymerase (EC 2.4.2.30) are detected within 15 min of detachment, 30-45 min before caspase 3 activation. Caspase 1 and caspase 10 are present as proenzymes, yet they remain inactive in response to this trigger of apoptosis. Human IEC are primed to rapidly undergo detachment-induced apoptosis involving the selective and sequential activation of preformed caspases. This study may enhance our understanding of physiological events occurring as IEC are shed. Their rapid apoptotic response to detachment may facilitate the high turnover of cells and ensure homeostasis in the intestinal epithelium.
Department of Medicine, School of Medicine, Case Western Reserve University, Cleveland, Ohio 44106-4952, USA.
J
J
Grossmann
S
S
Mohr
E G
EG
Lapentina
C
C
Fiocchi
A D
AD
Levine
D017209:Apoptosis; D020170:Caspase 1; D053148:Caspase 3; D053178:Caspase 6; D020169:Caspases; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D004789:Enzyme Activation; D004847:Epithelial Cells / Q000201:enzymology; D006801:Humans; D007422:Intestines / Q000201:enzymology*; D007700:Kinetics; D034882:Lamins; D009687:Nuclear Proteins / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism
D034882:Lamins; D009687:Nuclear Proteins; D011065:Poly(ADP-ribose) Polymerases; C505407:CASP3 protein, human; C505514:CASP6 protein, human; D053148:Caspase 3; D053178:Caspase 6; D020169:Caspases; D003546:Cysteine Endopeptidases; D020170:Caspase 1