Virus-induced modification of the host cell is required for expression of the bacterial chloramphenicol acetyltransferase gene controlled by a late herpes simplex virus promoter (VP5).
Virus-induced modification of the host cell is required for expression of the bacterial chloramphenicol acetyltransferase gene controlled by a late herpes simplex virus promoter (VP5).
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病毒诱导的宿主细胞修饰是受晚期单纯疱疹病毒启动子 (VP5) 控制的细菌氯霉素乙酰转移酶基因表达所必需的。
DOI：
10.1128/jvi.56.1.19-30.1985
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发表时间：
1985
影响因子：
5.4
通讯作者：
Wagner,EK
中科院分区：
文献类型：
--
作者：
Costa,RH;Draper,KG;Devi-Rao,G;Thompson,RL;Wagner,EK
以细菌氯霉素乙酰转移酶基因为表达标记物，采用瞬时表达法检测了单纯疱疹病毒1型（HSV-1）早期（碱性外切酶）和晚期（VP5）基因启动子调控下基因的表达要求。这两种启动子都被诱导，导致在HSV-1低倍数感染时产生高水平的酶。从含有HSV-1启动子结构的感染细胞中分离的RNA与标记基因DNA杂交的S1核酸酶分析表明，在病毒启动子的引导下，标记基因的转录起始正常。病毒DNA序列不超过125个碱基5'的假定转录帽位点就足以使晚期启动子的活性达到最大。与早期基因控制的表达相反，晚期启动子在未感染细胞中没有可测量水平的活性，直到转录帽位点的75和125碱基5'之间的DNA序列被删除。用HSV启动子控制的表达标记和含有HSV α（即早期）基因的cosmid共转染细胞表明，早期启动子和晚期启动子的完全表达需要相同的病毒诱导的宿主细胞修饰。与正常病毒感染的情况相比，抑制病毒DNA合成导致在早期或晚期启动子控制下标记基因的瞬时表达率增加。这些数据提供了证据，证明晚期HSV基因的正常表达过程涉及感染细胞中潜在活性启动子的负调节。
The requirements for expression of genes under the control of early (alkaline exonuclease) and late (VP5) herpes simplex virus type 1 (HSV-1) gene promoters were examined in a transient expression assay, using the bacterial chloramphenicol acetyltransferase gene as an expression marker. Both promoters were induced, resulting in the production of high levels of the enzyme upon low-multiplicity infection by HSV-1. S1 nuclease analysis of hybrids between RNA isolated from infected cells containing HSV-1 promoter constructs and marker gene DNA demonstrated normal transcriptional initiation of the marker gene directed by the viral promoters. Viral DNA sequences no more than 125 bases 5' of the putative transcriptional cap site were sufficient for maximum activity of the late promoter. In contrast to expression controlled by the early gene, the late promoter was not active at a measurable level in uninfected cells until DNA sequences between 75 and 125 bases 5' of the transcriptional cap site were deleted. Cotransfection of cells with the expression marker controlled by HSV promoters and a cosmid containing HSV alpha (immediate-early) genes indicated that full expression of both early and late promoters requires the same virus-induced host cell modifications. Inhibition of viral DNA synthesis results in an increased rate of transient expression of marker genes under control of either early or late promoters in contrast to the situation in normal virus infection. These data provide evidence that the normal course of expression of late HSV genes involves negative modulation of potentially active promoters in the infected cell.