Activation of the virus-induced IKK/NF-κB signalling axis is critical for the replication of human cytomegalovirus in quiescent cells
Activation of the virus-induced IKK/NF-κB signalling axis is critical for the replication of human cytomegalovirus in quiescent cells
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DOI：
10.1111/j.1462-5822.2007.00936.x
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发表时间：
2007-08-01
影响因子：
3.4
通讯作者：
Gribaudo, Giorgio
中科院分区：
文献类型：
--
作者：
Caposio, Patrizia;Luganini, Anna;Gribaudo, Giorgio
IKK/ nf - κ B信号通路的激活是人巨细胞病毒（HCMV）感染的一个标志。然而，它在调节主要的立即早期启动子（MIEP）依赖性转录和HCMV复制中的作用仍然存在争议。本研究使用遗传方法的组合来研究细胞培养条件对内皮细胞或成纤维细胞感染期间病毒诱导的NF-kappa B激活的重要性的影响。在感染HCMV临床分离物后，腺病毒介导的IKK2激酶显性阴性突变体（dnIKK2）在人脐静脉内皮细胞中的表达导致I κ B α降解和nf - κ B活化的强烈降低。在感染前生长停滞的表达dnikk2的细胞中，病毒复制受损，而在复制细胞中则没有。dnIKK2的抑制作用与病毒株和使用的细胞类型无关，因为实验室AD169株在表达dnIKK2的静止成纤维细胞中复制也受到损害。此外，在临床分离的重组HCMV病毒中，MIEP内NF-kappa B应答元件的进行性破坏阻止了它们在静止细胞中的复制，而不是在活跃生长的细胞中复制。这些结果表明，病毒诱导的IKK/NF-kappa B活性在静止细胞中触发病毒IE基因表达和生产性复制方面发挥了重要作用。
Activation of the IKK/NF-kappa B signalling pathway is a hallmark of human cytomegalovirus (HCMV) infection. However, its role in regulating major immediate-early promoter (MIEP)-dependent transcription and HCMV replication remains controversial. This study uses a combination of genetic approaches to investigate the effects of cell culture conditions on the importance of virus-induced NF-kappa B activation during the infection of endothelial cells or fibroblasts. Adenoviral-mediated expression of a dominant-negative mutant of IKK2 kinase (dnIKK2) in human umbilical vein endothelial cells resulted in a strong reduction of I kappa B alpha degradation and NF-kappa B activation following infection with an HCMV clinical isolate. Viral replication was impaired in dnIKK2-expressing cells that were growth-arrested before infection, but not in replicating cells. The inhibitory effect of dnIKK2 was independent from the virus strain and the cell type used, because replication of the laboratory AD169 strain was impaired as well in dnIKK2-expressing quiescent fibroblasts. Moreover, progressive disruption of NF-kappa B response elements within the MIEP in recombinant HCMV viruses derived from the clinical isolate prevented their replication in quiescent cells but not in actively growing cells. These results demonstrate an essential role of virus-induced IKK/NF-kappa B activity to trigger both viral IE gene expression and productive replication in quiescent cells.