Executioner caspase-3,-6, and-7 perform distinct, non-redundant roles during the demolition phase of apoptosis
Executioner caspase-3,-6, and-7 perform distinct, non-redundant roles during the demolition phase of apoptosis
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DOI：
10.1074/jbc.m008363200
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发表时间：
2001-03-09
影响因子：
4.8
通讯作者：
Martin, SJ
中科院分区：
文献类型：
--
作者：
Slee, EA;Adrain, C;Martin, SJ
细胞凋亡是由半胱氨酸蛋白酶家族调控的：半胱氨酸蛋白酶。已鉴定出14种哺乳动物caspase，其中3种(caspase-3、-6和-7)被认为通过裂解多种结构蛋白和修复蛋白来协调细胞凋亡的执行阶段。然而，“刽子手”半胱氨酸天冬氨酸酶对细胞破坏的相对贡献：仍然是推测的。在这里，我们使用了免疫耗尽的caspase-3、-6或-7的无细胞提取物来检测14种caspase底物的凋亡相关蛋白分解以及核凝聚、染色质边集和DNA片段化对caspase的要求。我们发现caspase-3是caspase的主要执行者，是细胞色素c/dATP诱导的对Fodrin、明胶蛋白、U1小核糖核蛋白、DNA碎片因子45(DFF45)/caspase激活的DNA酶抑制因子(ICAD)、受体相互作用蛋白(RIP)、X连锁的凋亡抑制蛋白(X-IAP)、信号转导和转录激活因子-1(STAT1)、拓扑异构酶I、波形蛋白、Rb和lamin B所必需的，但对多聚核糖聚合酶(PARP)或lamin A的切割不是必需的。此外，caspase-3对与细胞凋亡相关的染色质边集、染色质边集、波形蛋白、Rb和lamin B的切割也是必不可少的。DNA碎裂，以及这个系统中的核崩溃。令人惊讶的是，尽管caspase-6和-7被认为是重要的下游效应caspase，但这两个caspase的缺失对所研究的任何参数的影响都很小，这使得人们对它们在细胞凋亡执行阶段的确切作用产生了怀疑。
Apoptosis is orchestrated by a family of cysteine proteases known: as the caspases. Fourteen mammalian caspases have been identified, three of which (caspase-3, -6, and -7) are thought to coordinate the execution phase of apoptosis by cleaving multiple structural and repair proteins. However, the relative contributions that the "executioner" caspases make to the demolition of the cell: remains speculative. Here we have used cell-free extracts immune-depleted of either caspase-3, -6, or -7 to examine the caspase requirements for apoptosis-associated proteolysis of 14 caspase substrates as well as nuclear condensation, chromatin margination, and DNA fragmentation. We show that caspase-3 is the primary executioner caspase in this system, necessary for cytochrome c/dATP-inducible cleavage of fodrin, gelsolin, U1 small nuclear ribonucleopretein, DNA fragmentation factor 45 (DFF45)/inhibitor of caspase-activated DNase (ICAD), receptor-interacting protein (RIP), X-linked inhibitor of apoptosis protein (X-IAP), signal transducer and activator of transcription-1 (STAT1), topoisomerase I, vimentin, Rb, and lamin B but not for cleavage of poly(ADP-ribose) polymerase (PARP) or lamin A. In addition, caspase-3 was also essential for apoptosis-associated: chromatin margination, DNA fragmentation, and nuclear collapse in this system. Surprisingly, although caspase-6 and -7 are considered to be important downstream-effector caspases, depletion of either caspase had minimal impact on any of the parameters investigated, calling into question their precise role during the execution phase of apoptosis.