Cell type-specific cleavage of nucleocapsid protein by effector caspases during SARS coronavirus infection.
Cell type-specific cleavage of nucleocapsid protein by effector caspases during SARS coronavirus infection.
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在SARS冠状病毒感染期间，通过效应子胱天蛋白酶对细胞类型的特异性裂解。
DOI：
10.1016/j.jmb.2007.11.081
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发表时间：
2008-02-08
影响因子：
5.6
通讯作者：
Gilch S
中科院分区：
文献类型：
--
作者：
Diemer C;Schneider M;Seebach J;Quaas J;Frösner G;Schätzl HM;Gilch S
2003年爆发的严重急性呼吸综合征（SARS）疫情是由一种新型冠状病毒（CoV）引起的，命名为SARS-CoV。SARS冠状病毒的RNA基因组与核衣壳蛋白（N）复合形成螺旋状核衣壳。除了这一主要功能，N似乎参与凋亡的情况。我们表明，在感染Vero E6细胞与SARS-CoV，这elevens一个显着的细胞病变效应和高病毒滴度，N被切割的半胱天冬酶。相反，在SARS-CoV感染的Caco-2细胞中，显示出中度的细胞病变效应和低病毒滴度，没有观察到N的这种处理。为了进一步验证这些观察结果，我们在不同的细胞系中瞬时表达N。Caco-2和N2 a细胞作为SARS-CoV持续感染的模型，而Vero E6和A549细胞作为SARS-CoV裂解感染的原型细胞系。实验表明，N诱导的内在凋亡途径，导致在处理的N在残基400和403由半胱天冬酶-6和/或半胱天冬酶-3。值得注意的是，半胱天冬酶的激活在SARS冠状病毒感染以及瞬时转染的细胞中具有高度的细胞类型特异性。在Caco-2和N2 a细胞中，几乎没有检测到N-加工。在Vero E6和A549细胞中，高比例的N被半胱天冬酶切割。此外，我们研究了SARS-CoV N在这些细胞系中的亚细胞定位。在转染的Vero E6和A549细胞中，SARS-CoV N定位于细胞质和细胞核，而在Caco-2和N2 a细胞中，几乎没有观察到核定位。此外，我们的研究表明，核定位的N是必不可少的caspase-6介导的切割。这些数据表明SARS-CoV的复制周期、N的亚细胞定位、细胞凋亡的诱导以及随后导致N切割的半胱天冬酶激活之间存在相关性。
The epidemic outbreak of severe acute respiratory syndrome (SARS) in 2003 was caused by a novel coronavirus (CoV), designated SARS-CoV. The RNA genome of SARS-CoV is complexed by the nucleocapsid protein (N) to form a helical nucleocapsid. Besides this primary function, N seems to be involved in apoptotic scenarios. We show that upon infection of Vero E6 cells with SARS-CoV, which elicits a pronounced cytopathic effect and a high viral titer, N is cleaved by caspases. In contrast, in SARS-CoV-infected Caco-2 cells, which show a moderate cytopathic effect and a low viral titer, this processing of N was not observed. To further verify these observations, we transiently expressed N in different cell lines. Caco-2 and N2a cells served as models for persistent SARS-CoV infection, whereas Vero E6 and A549 cells did as prototype cell lines lytically infected by SARS-CoV. The experiments revealed that N induces the intrinsic apoptotic pathway, resulting in processing of N at residues 400 and 403 by caspase-6 and/or caspase-3. Of note, caspase activation is highly cell type specific in SARS-CoV-infected as well as transiently transfected cells. In Caco-2 and N2a cells, almost no N-processing was detectable. In Vero E6 and A549 cells, a high proportion of N was cleaved by caspases. Moreover, we examined the subcellular localization of SARS-CoV N in these cell lines. In transfected Vero E6 and A549 cells, SARS-CoV N was localized both in the cytoplasm and nucleus, whereas in Caco-2 and N2a cells, nearly no nuclear localization was observed. In addition, our studies indicate that the nuclear localization of N is essential for its caspase-6-mediated cleavage. These data suggest a correlation among the replication cycle of SARS-CoV, subcellular localization of N, induction of apoptosis, and the subsequent activation of caspases leading to cleavage of N.
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DOI：
10.1016/j.bbrc.2004.05.107
发表时间：
2004-07-09
影响因子：
3.1
作者：
Mizutani T;Fukushi S;Saijo M;Kurane I;Morikawa S
通讯作者：
Morikawa S
影响因子：
64.8
作者：
Li W;Moore MJ;Vasilieva N;Sui J;Wong SK;Berne MA;Somasundaran M;Sullivan JL;Luzuriaga K;Greenough TC;Choe H;Farzan M
通讯作者：
Farzan M
影响因子：
3.7
作者：
Liao, QJ;Ye, LB;Wu, ZH
通讯作者：
Wu, ZH
影响因子：
3.8
作者：
Law, PTW;Wong, CH;Tsui, SKW
通讯作者：
Tsui, SKW
影响因子：
5.4
作者：
Kopecky-Bromberg, Sarah A.;Martinez-Sobrido, Luis;Palese, Peter
通讯作者：
Palese, Peter