EHV-1 的 UL4 蛋白调节病毒基因表达，与细胞转录因子相互作用。
The EHV-1 UL4 protein that tempers viral gene expression interacts with cellular transcription factors.
机构信息
Center for Molecular and Tumor Virology, Department of Microbiology and Immunology, Louisiana State University Health Sciences Center, 1501 Kings Highway, Shreveport, LA 71130-3932, USA.
Center for Molecular and Tumor Virology, Department of Microbiology and Immunology, Louisiana State University Health Sciences Center, 1501 Kings Highway, Shreveport, LA 71130-3932, USA.
出版信息
Virology. 2014 Jan 20;449:25-34. doi: 10.1016/j.virol.2013.11.005. Epub 2013 Nov 21.
The UL4 gene is conserved within the genome of defective interfering particles of equine herpesvirus type 1 (EHV-1) that mediate persistent infection. Here, we show that the UL4 protein inhibits EHV-1 reporter gene expression by decreasing the level of transcribed mRNA. The UL4 protein did not bind any gene class of EHV-1 promoters in electromobility or chromatin immunoprecipitation assays, but directly interacted with the TATA box-binding protein (TBP) and the carboxy-terminal domain of RNA polymerase II both in vitro (GST-pulldown assays) and in infected cells (coimmunoprecipitation analyses). Microarray analyses of the expression of the 78 EHV-1 genes revealed that viral late genes important for virion assembly displayed enhanced expression in cells infected with UL4-null virus as compared to wild-type or UL4-restored EHV-1. Quantitative PCR analyses showed that viral DNA replication was not retarded in cells infected with the UL4-null virus as compared to wild-type EHV-1.
UL4 基因在介导持续性感染的马疱疹病毒 1 型（EHV-1）缺陷干扰颗粒的基因组中是保守的。在这里，我们表明 UL4 蛋白通过降低转录 mRNA 的水平来抑制 EHV-1 报告基因的表达。UL4 蛋白在电泳或染色质免疫沉淀分析中没有与任何一类 EHV-1 启动子结合，但在体外（GST 下拉分析）和感染细胞中（共免疫沉淀分析）直接与 TATA 框结合蛋白（TBP）和 RNA 聚合酶 II 的羧基末端结构域相互作用。EHV-1 的 78 个基因表达的微阵列分析显示，对于病毒粒子组装很重要的病毒晚期基因在感染 UL4 缺失病毒的细胞中与野生型或 UL4 恢复的 EHV-1 相比表达增强。定量 PCR 分析表明，与野生型 EHV-1 相比，在感染 UL4 缺失病毒的细胞中，病毒 DNA 复制并没有受到阻碍。
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