Reovirus-induced apoptosis requires activation of transcription factor NF-κB
Reovirus-induced apoptosis requires activation of transcription factor NF-κB
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DOI：
10.1128/jvi.74.7.2981-2989.2000
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发表时间：
2000-04-01
影响因子：
5.4
通讯作者：
Dermody, TS
中科院分区：
文献类型：
--
作者：
Connolly, JL;Rodgers, SE;Dermody, TS
呼肠孤病毒感染诱导培养细胞和体内细胞凋亡。为了确定介导这种反应的宿主细胞因子，我们研究了呼肠孤病毒感染是否改变了转录因子核因子κ B（NF-κ B）的激活状态。如在电泳迁移率变动测定中所确定的，HeLa细胞的呼肠孤病毒感染导致含有Rel家族成员p50和p65的NF-κ B复合物的核转位。呼肠孤病毒诱导的NF-κ B B DNA结合活性的激活与报告基因测定中NF-κ B定向转录的开始相关。三个独立的证据表明，这种功能形式的NF-κ B是呼肠孤病毒诱导的细胞凋亡所必需的。首先，用蛋白酶体抑制剂处理呼肠孤病毒感染的HeLa细胞可防止感染后NF-κ B活化，并显著减少呼肠孤病毒诱导的细胞凋亡。第二，瞬时表达I κ B的显性负性形式，组成性抑制NF-κ B活化，显著降低呼肠孤病毒感染引发的细胞凋亡水平。第三，与表达完整NF-κ B信号通路的细胞相比，NF-κ B B p50或p65亚基缺陷的突变细胞系对呼肠孤病毒诱导的细胞凋亡具有抗性。这些发现表明NF-κ B在呼肠孤病毒诱导易感宿主细胞凋亡的机制中起重要作用。
Reovirus infection induces apoptosis in cultured cells and in vivo. To identify host cell factors that mediate this response, we investigated whether reovirus infection alters the activation state of the transcription factor nuclear factor kappa B (NF-kappa B). As determined in electrophoretic mobility shift assays, reovirus infection of HeLa cells leads to nuclear translocation of NF-kappa B complexes containing Rel family members p50 and p65. Reovirus induced activation of NF-kappa B DNA-binding activity correlated with the onset of NF-kappa B-directed transcription in reporter gene assays. Three independent lines of evidence indicate that this functional form of NF-kappa B is required for reovirus-induced apoptosis. First, treatment of reovirus-infected HeLa cells with a proteasome inhibitor prevents NF-kappa B activation following infection and substantially diminishes reovirus-induced apoptosis. Second, transient expression of a dominant-negative form of I kappa B that constitutively represses NF-kappa B activation significantly reduces levels of apoptosis triggered by reovirus infection. Third, mutant cell lines deficient for either the p50 or p65 subunits of NF-kappa B are resistant to reovirus-induced apoptosis compared with cells expressing an intact NF-kappa B signaling pathway. These findings indicate that NF-kappa B plays a significant role in the mechanism by which reovirus induces apoptosis in susceptible host cells.