Everett R D
MRC Virology Unit, Glasgow, UK.
EMBO J. 1987 Jul;6(7):2069-76. doi: 10.1002/j.1460-2075.1987.tb02472.x.
Viral genes expressed during infection of tissue culture cells by herpes simplex virus type 1 (HSV-1) can be divided into three broad temporal groups called immediate-early (IE), early and late. Prior expression of IE gene products is required before later classes of genes can be transcribed. Using cloned copies of viral genes in plasmid vectors, it has been shown that the products of IE genes 1 and 3 (Vmw110 and Vmw175) can activate transcription from both viral and cellular promoters in short-term transfection assays. The effects of these two IE gene products are markedly synergistic in HeLa cells; the activation observed with both genes present is about 20 times that observed with Vmw175 alone. The mechanism of this activation and the properties of the IE proteins that are required are not well understood. This paper describes the construction of a large number of in-frame insertion and deletion mutations in a plasmid-encoded copy of IE gene 1. The ability of the mutant Vmw110 polypeptides to activate gene expression (in the presence of Vmw175) from the HSV-1 glycoprotein gD promoter linked to the chloramphenicol acetyl transferase gene was studied. The results show that the structural integrity of at least five regions of the polypeptide are important for its function in the presence of Vmw175.
单纯疱疹病毒1型(HSV-1)感染组织培养细胞期间表达的病毒基因可分为三个大致的时间组,即立即早期(IE)、早期和晚期。在转录后期基因类别之前,需要先表达IE基因产物。利用质粒载体中病毒基因的克隆拷贝,已表明IE基因1和3的产物(Vmw110和Vmw175)在短期转染试验中可激活病毒和细胞启动子的转录。在HeLa细胞中,这两种IE基因产物的作用具有明显的协同性;同时存在这两个基因时观察到的激活作用约为单独使用Vmw175时的20倍。这种激活的机制以及所需的IE蛋白的特性尚不清楚。本文描述了在IE基因1的质粒编码拷贝中构建大量框内插入和缺失突变体的过程。研究了突变的Vmw110多肽在存在Vmw175的情况下激活与氯霉素乙酰转移酶基因相连的HSV-1糖蛋白gD启动子的基因表达的能力。结果表明,该多肽至少五个区域的结构完整性对其在Vmw175存在时的功能很重要。