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首页> 美国卫生研究院文献>Journal of Virology >Transcription of Herpes Simplex Virus Immediate-Early and Early Genes Is Inhibited by Roscovitine an Inhibitor Specific for Cellular Cyclin-Dependent Kinases
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Transcription of Herpes Simplex Virus Immediate-Early and Early Genes Is Inhibited by Roscovitine an Inhibitor Specific for Cellular Cyclin-Dependent Kinases

机译：Roscovitine抑制单纯疱疹病毒的早期和早期基因转录Roscovitine是细胞周期蛋白依赖性激酶特异的抑制剂。

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摘要

Although herpes simplex virus (HSV) replicates in noncycling as well as cycling cells, including terminally differentiated neurons, it has recently been shown that viral replication requires the activities of cellular cyclin-dependent kinases (cdks) (L. M. Schang, J. Phillips, and P. A. Schaffer, J. Virol. 72:5626–5637, 1998). Since we were unable to isolate HSV mutants resistant to two cdk inhibitors, Olomoucine and Roscovitine (Rosco), we hypothesized that cdks may be required for more than one viral function during HSV replication. In the experiments presented here, we tested this hypothesis by measuring the efficiency of (i) viral replication; (ii) expression of selected immediate-early (IE) (ICP0 and ICP4), early (E) (ICP8 and TK), and late (L) (gC) genes; and (iii) viral DNA synthesis in infected cultures to which Rosco was added after IE or IE and E proteins had already been synthesized. Rosco inhibited HSV replication, transcription of IE and E genes, and viral DNA synthesis when added at 1, 2, or 6 h postinfection or after release from a 6-h cycloheximide block. Transcription of a representative L gene, gC, was also inhibited by Rosco under all conditions examined. We conclude from these studies that cellular cdks are required for transcription of E as well as IE genes. In contrast, steady-state levels of at least one cellular housekeeping gene were not affected by Rosco. The requirement of viral IE and E transcription for cellular cdks may reflect either a requirement for specific cdk-activated cellular and/or viral transcription factors or a more global requirement for cdks in the transcriptional activation of the viral genome.
机译：尽管单纯疱疹病毒（HSV）在非循环以及循环细胞（包括终末分化的神经元）中复制，但最近发现病毒复制需要细胞周期蛋白依赖性激酶（cdks）的活性（LM Schang，J.Phillips和PA Schaffer，J. Virol。72：5626-5637，1998）。由于我们无法分离出对两种cdk抑制剂Olomoucine和Roscovitine（Rosco）有抵抗力的HSV突变体，因此我们假设在HSV复制过程中，cdks可能需要一种以上的病毒功能。在这里介绍的实验中，我们通过测量（i）病毒复制的效率来检验这个假设。 （ii）选定的即早（IE）（ICP0和ICP4），早期（E）（ICP8和TK）和晚期（L）（gC）基因的表达； （iii）在已经合成IE或IE和E蛋白之后在其中添加了Rosco的感染培养物中的病毒DNA合成。当在感染后1、2或6小时或从6小时的环己酰亚胺嵌段中释放后，Rosco抑制了HSV复制，IE和E基因的转录以及病毒DNA的合成。在所有检查条件下，Rosco也抑制了代表性L基因gC的转录。从这些研究中我们得出结论，细胞cdks是E和IE基因转录所必需的。相比之下，至少一种细胞管家基因的稳态水平不受Rosco的影响。细胞cdks对病毒IE和E转录的需求可能反映了对特定cdk活化的细胞和/或病毒转录因子的需求，或者反映了病毒基因组转录激活中对cdks的更全面的需求。

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