Temporal association of the herpes simplex virus genome with histone proteins during a lytic infection
Temporal association of the herpes simplex virus genome with histone proteins during a lytic infection
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DOI：
10.1128/jvi.00586-07
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发表时间：
2008-04-01
影响因子：
5.4
通讯作者：
Fraser, Nigel W.
中科院分区：
文献类型：
--
作者：
Oh, Jaewook;Fraser, Nigel W.
以前的工作已经确定，在裂解感染期间，单纯疱疹病毒（HSV）基因组上存在核小体，但它们并不像细胞DNA那样以等间距排列。然而，像在细胞DNA中一样，几种病毒基因的启动子区已显示与含有修饰的组蛋白的核小体相关，所述修饰的组蛋白通常被发现与活跃转录的基因相关。此外，已经发现，经修饰的组蛋白与HSV基因组的关联可以在感染后的最早时间（感染后1小时）检测到，并增加至感染后3小时。然而，从3小时至6小时感染后（复制周期的后期），协会下降。在这项研究中，我们研究了组蛋白与所有动力学类基因的启动子区域的关联。这是在Sy5y细胞中感染的时间过程中使用蔗糖梯度沉降、溴脱氧尿苷标记、染色质免疫沉淀测定、蛋白质印迹分析、胰蛋白酶和DNA酶消化以及定量实时PCR来完成的。因为在HSV 1型衣壳内没有检测到组蛋白，所以病毒基因组可能在从感染病毒体转运到宿主细胞核后开始与组蛋白结合。所有基因类别的启动子区（即早期、早期和晚期）在病毒基因表达开始时与组蛋白结合。然而，在病毒DNA复制启动后，组蛋白似乎不与新合成的病毒基因组相关。
Previous work has determined that there are nucleosomes on the herpes simplex virus (HSV) genome during a lytic infection but that they are not arranged in an equally spaced array like in cellular DNA. However, like in cellular DNA, the promoter regions of several viral genes have been shown to be associated with nucleosomes containing modified histone proteins that are generally found associated with actively transcribed genes. Furthermore, it has been found that the association of modified histones with the HSV genome can be detected at the earliest times postinfection (1 h postinfection) and increases up to 3 h postinfection. However from 3 h to 6 h postinfection (the late phase of the replication cycle), the association decreases. In this study we have examined histone association with promoter regions of all kinetic classes of genes. This was done over the time course of an infection in Sy5y cells using sucrose gradient sedimentation, bromodeoxyuridine labeling, chromatin immunoprecipitation assays, Western blot analysis, trypsin and DNase digestion, and quantitative real-time PCR. Because no histones were detected inside HSV type 1 capsids, the viral genome probably starts to associate with histones after being transported from infecting virions into the host nucleus. Promoter regions of all gene classes (immediate early, early, and late) bind with histone proteins at the start of viral gene expression. However, after viral DNA replication initiates, histones appear not to associate with newly synthesized viral genomes.