TY - JOUR
T1 - Apopain/CPP32 cleaves proteins that are essential for cellular repair
T2 - A fundamental principle of apoptotic death
AU - Casciola-Rosen, Livia
AU - Nicholson, Donald W.
AU - Chong, Tae
AU - Rowan, Kevin R.
AU - Thornberry, Nancy A.
AU - Miller, Douglas K.
AU - Rosen, Antony
PY - 1996/5/1
Y1 - 1996/5/1
N2 - Proteolysis mediated by the interleukin 1β-converting enzyme (ICE) homologues is an important mechanism of the apoptotic process. The ICE homologue apopain/CPP-32/Yama (subsequently referred to as apopain) cleaves poly(ADP-ribose)polymerase (PARP) early during apoptosis. Additional apoptosis-specific protein cleavages have been observed in which the direct involvement of ICE-like proteases has been postulated. These substrates include the 70-kD protein component of the U1-ribonucleoprotein (U1-70kD), and the catalytic subunit of the DNA-dependent protein kinase (DNA-PK(cs)). The present studies demonstrate that U1-70kD and DNA-PK(cs) are excellent substrates for apopain, with cleavage occurring at sites that are highly similar to the cleavage site within PARP. The fragments generated from isolated protein substrates by apopain are identical to those observed in intact apoptotic cells, in apoptotic cell extracts, and in normal cell extracts to which apopain has been added. Like PARP, cleavage of these substrates in apoptotic cell extracts is abolished by nanomolar concentrations of Ac-DEVD-CHO and micromolar amounts of Ac-YVAD-CHO, confirming the involvement of apopain or an apopain-like activity. We propose that a central function of apopain or similar homologues in apoptosis is the cleavage of nuclear repair proteins, thereby abolishing their critical homeostatic functions.
AB - Proteolysis mediated by the interleukin 1β-converting enzyme (ICE) homologues is an important mechanism of the apoptotic process. The ICE homologue apopain/CPP-32/Yama (subsequently referred to as apopain) cleaves poly(ADP-ribose)polymerase (PARP) early during apoptosis. Additional apoptosis-specific protein cleavages have been observed in which the direct involvement of ICE-like proteases has been postulated. These substrates include the 70-kD protein component of the U1-ribonucleoprotein (U1-70kD), and the catalytic subunit of the DNA-dependent protein kinase (DNA-PK(cs)). The present studies demonstrate that U1-70kD and DNA-PK(cs) are excellent substrates for apopain, with cleavage occurring at sites that are highly similar to the cleavage site within PARP. The fragments generated from isolated protein substrates by apopain are identical to those observed in intact apoptotic cells, in apoptotic cell extracts, and in normal cell extracts to which apopain has been added. Like PARP, cleavage of these substrates in apoptotic cell extracts is abolished by nanomolar concentrations of Ac-DEVD-CHO and micromolar amounts of Ac-YVAD-CHO, confirming the involvement of apopain or an apopain-like activity. We propose that a central function of apopain or similar homologues in apoptosis is the cleavage of nuclear repair proteins, thereby abolishing their critical homeostatic functions.
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U2 - 10.1084/jem.183.5.1957
DO - 10.1084/jem.183.5.1957
M3 - Article
C2 - 8642305
AN - SCOPUS:0029890823
SN - 0022-1007
VL - 183
SP - 1957
EP - 1964
JO - Journal of Experimental Medicine
JF - Journal of Experimental Medicine
IS - 5
ER -