Efficient replication by herpes simplex virus type 1 involves activation of the IκB kinase-IκB-p65 pathway
Efficient replication by herpes simplex virus type 1 involves activation of the IκB kinase-IκB-p65 pathway
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DOI：
10.1128/jvi.78.24.13582-13590.2004
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发表时间：
2004-12-01
影响因子：
5.4
通讯作者：
Bachenheimer, SL
中科院分区：
文献类型：
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作者：
Gregory, D;Hargett, D;Bachenheimer, SL
单纯疱疹病毒1型(HSV-1)感染可诱导NFkappaB的持续性核转位。为了确定NFkappaB的上游效应因子及其对病毒复制的影响，我们使用了缺失IKK1或IKK2的小鼠胚胎成纤维细胞(MEF)来源的细胞系，IKK1或IKK2是IkappaB激酶(IKK)复合体的催化亚单位。检测感染的MEF的病毒产量、IkappaBalpha丢失、p65核转位和NFkappaB DNA结合活性。与正常的MEF相比，缺乏IKK1或IKK2导致病毒产量损失86%~94%，IkappaBalpha损失很少或没有损失，并极大地减少了NFkappaB核转位。与病毒产量降低一致，晚期蛋白VP16和GC的积累受到严重抑制。用表达显性阴性(DN)IkappaBalpha的腺病毒载体感染正常的MEF、Hep2或A549细胞，然后重叠感染HSV，导致病毒产量下降98%。这些结果表明，IKK-IkappaB-P65途径在病毒感染后激活NFkappaB。对对照和双链RNA激活蛋白激酶缺失的MEF中NFkappaB的激活和病毒复制的分析表明，NFkappaB在NFkappaB的激活途径中没有作用。最后，在NFkappaB因DNIkappaB表达而被阻断的细胞中，HSV未能抑制细胞表面Annexin V染色和PARP裂解这两个凋亡标志物。这些结果支持一种模型，在该模型中，NFkappaB的激活至少部分通过抑制宿主对病毒感染的固有反应来促进HSV的有效复制。
Infection by herpes simplex virus type 1 (HSV-1) induces a persistent nuclear translocation of NFkappaB. To identify upstream effectors of NFkappaB and their effect on virus replication, we employed mouse embryo fibroblast (MEF)-derived cell lines with deletions of either IKK1 or IKK2, the catalytic subunits of the IkappaB kinase (IKK) complex. Infected MEFs were assayed for virus yield, loss of IkappaBalpha, nuclear translocation of p65, and NFkappaB DNA-binding activity. Absence of either IKK1 or IKK2 resulted in an 86 to 94% loss of virus yield compared to that of normal MEFs, little or no loss of IkappaBalpha, and greatly reduced NFkappaB nuclear translocation. Consistent with reduced virus yield, accumulation of the late proteins VP16 and gC was severely depressed. Infection of normal MEFs, Hep2, or A549 cells with an adenovirus vector expressing a dominant-negative (DN) IkappaBalpha, followed by superinfection with HSV, resulted in a 98% drop in virus yield. These results indicate that the IKK-IkappaB-p65 pathway activates NFkappaB after virus infection. Analysis of NFkappaB activation and virus replication in control and double-stranded RNA-activated protein kinase-null MEFs indicated that this kinase plays no role in the NFkappaB activation pathway. Finally, in cells where NFkappaB was blocked because of DNIkappaB expression, HSV failed to suppress two markers of apoptosis, cell surface Annexin V staining and PARP cleavage. These results support a model in which activation of NFkappaB promotes efficient replication by HSV, at least in part by suppressing a host innate response to virus infection.