Caspase activation and specific cleavage of substrates after coxsackievirus B3-induced cytopathic effect in HeLa cells
Caspase activation and specific cleavage of substrates after coxsackievirus B3-induced cytopathic effect in HeLa cells
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DOI：
10.1128/jvi.72.9.7669-7675.1998
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发表时间：
1998-09-01
影响因子：
5.4
通讯作者：
McManus, BM
中科院分区：
文献类型：
--
作者：
Carthy, CM;Granville, DJ;McManus, BM
柯萨奇病毒B3（Coxsackievirus B3，CVB 3）是小核糖核酸病毒科的一种肠道病毒，在感染后早期可引起细胞病变，并直接损伤体内多个易感器官和组织，包括心肌。CVB 3感染的HeLa细胞中细胞死亡途径的生化分析表明，在感染的HeLa细胞中观察到的退行性形态学变化之后，半胱天冬酶3的32-kDa预制体被裂解。半胱天冬酶活化测定证实，裂解的半胱天冬酶3是蛋白水解活性的。caspase 3底物聚（ADP-核糖）聚合酶，DNA修复酶，和DNA片段化因子，负责DNA片段化的核酸内切酶的细胞质抑制剂，在感染后9小时降解，产生其特征性的切割片段。用苄氧羰基-Val-Ala-Asp-fluoromethylketone（ZVAD. fetamine）抑制半胱天冬酶的激活并不能抑制病毒诱导的细胞病变效应，而在用卟啉光敏剂苯并卟啉衍生物单酸环A和可见光处理诱导的对照凋亡细胞中，ZVAD. fetamine抑制半胱天冬酶的激活则抑制凋亡表型。半胱天冬酶的激活和底物的裂解可能不是由小RNA病毒感染产生的特征性细胞病变效应的原因，但可能与细胞稳态过程和结构完整性的后期改变有关。
Coxsackievirus B3 (CVB3), an enterovirus in the family Picornaviridae, induces cytopathic changes in cell culture systems and directly injures multiple susceptible organs and tissues in vivo, including the myocardium, early after infection. Biochemical analysis of the cell death pathway in CVB3-infected HeLa cells demonstrated that the 32-kDa preform of caspase 3 is cleaved subsequent to the degenerative morphological changes seen in infected HeLa cells. Caspase activation assays confirm that the cleaved caspase 3 is proteolytically active. The caspase 3 substrates poly(ADP-ribose) polymerase, a DNA repair enzyme, and DNA fragmentation factor, a cytoplasmic inhibitor of an endonuclease responsible for DNA fragmentation, were degraded at 9 h following infection, yielding their characteristic cleavage fragments. Inhibition of caspase activation by benzyloqcarbonyl-Val-Ala-Asp-fluoromethylketone (ZVAD.fmk) did not inhibit the virus-induced cytopathic effect, while inhibition of caspase activation by ZVAD.fmk in control apoptotic cells induced by treatment with the porphyrin photosensitizer benzoporphyrin derivative monoacid ring A and visible light inhibited the apoptotic phenotype. Caspase activation and cleavage of substrates may not be responsible for the characteristic cytopathic effect produced by picornavirus infection yet may be related to late-stage alterations of cellular homeostatic processes and structural integrity.