Two gamma interferon-activated site-like elements in the human cytomegalovirus major immediate-early promoter/enhancer are important for viral replication.
Two gamma interferon-activated site-like elements in the human cytomegalovirus major immediate-early promoter/enhancer are important for viral replication.
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人巨细胞病毒主要立即早期启动子/增强子中的两个γ干扰素激活的位点样元件对于病毒复制很重要。
DOI：
10.1128/jvi.79.8.5035-5046.2005
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Zhu,Hua
中科院分区：
文献类型：
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作者：
Netterwald,James;Yang,Shaojun;Wang,Weijia;Ghanny,Salena;Cody,Michael;Soteropoulos,Patricia;Tian,Bin;Dunn,Walter;Liu,Fenyong;Zhu,Hua
人巨细胞病毒（HCMV）感染直接启动信号转导途径，导致大量细胞干扰素刺激基因（ISG）的激活。我们以前的研究表明，两个干扰素反应元件，干扰素刺激的反应元件和γ干扰素激活位点（GAS），在ISG启动子作为HCMV反应位点（VRS）。有趣的是，两个GAS样VRS元件（VRS 1）也存在于HCMV主要立即早期启动子增强子（MIEP/E）中。在这项研究中，这些VRS元件在病毒复制的重要性进行了调查。我们证明，表达的主要IE基因，IE 1和IE 2，是干扰素诱导。为了理解该信号转导途径在HCMV主要IE表达中的生物学意义，突变MIEP/E中的两个VRS 1。VRS元件缺失或含有点突变的突变型HCMV在低感染复数（MOI）下比野生型病毒生长显著更慢。将野生型VRS 1插入突变病毒基因组中拯救了缓慢生长表型。此外，主要IE RNA和蛋白质的表达水平在低MOI下用VRS突变体感染期间大大降低。HCMV微阵列分析表明，VRS突变病毒感染宿主细胞导致病毒基因表达的整体减少。总的来说，这些数据表明MIEP/E中的两个VRS元件对于有效的病毒基因表达和复制是必需的。这项研究表明，虽然HCMV启动的信号转导途径导致诱导细胞的抗病毒基因，它也起到刺激病毒主要IE基因的表达。这可能是一种新的病毒策略，其中该途径用于调节基因表达并在再激活中发挥作用。
Human cytomegalovirus (HCMV) infection directly initiates a signal transduction pathway that leads to activation of a large number of cellular interferon-stimulated genes (ISGs). Our previous studies demonstrated that two interferon response elements, the interferon-stimulated response element and gamma interferon-activated site (GAS), in the ISG promoters serve as HCMV response sites (VRS). Interestingly, two GAS-like VRS elements (VRS1) were also present in the HCMV major immediate-early promoter-enhancer (MIEP/E). In this study, the importance of these VRS elements in viral replication was investigated. We demonstrate that the expression of the major IE genes, IE1 and IE2, is interferon inducible. To understand the biological significance of this signal transduction pathway in HCMV major IE expression, the two VRS1 in the MIEP/E were mutated. Mutant HCMVs in which the VRS elements were deleted or that contained point mutations grew dramatically more slowly than wild-type virus at a low multiplicity of infection (MOI). Insertion of wild-type VRS1 into the mutant viral genome rescued the slow growth phenotype. Furthermore, the expression levels of major IE RNAs and proteins were greatly reduced during infection with the VRS mutants at a low MOI. HCMV microarray analysis indicated that infection of host cells with the VRS mutant virus resulted in a global reduction in the expression of viral genes. Collectively, these data demonstrate that the two VRS elements in the MIEP/E are necessary for efficient viral gene expression and replication. This study suggests that although the HCMV-initiated signal transduction pathway results in induction of cellular antiviral genes, it also functions to stimulate viral major IE gene expression. This might be a new viral strategy in which the pathway is used to regulate gene expression and play a role in reactivation.