Ace C I, Dalrymple M A, Ramsay F H, Preston V G, Preston C M
Medical Research Council Virology Unit, Glasgow, U.K.
J Gen Virol. 1988 Oct;69 ( Pt 10):2595-605. doi: 10.1099/0022-1317-69-10-2595.
The herpes simplex virus type 1 (HSV-1) polypeptide Vmw65 is a structural component of the virus particle and is also responsible for trans-induction of immediate early (IE) transcription. Functional domains of this polypeptide were investigated by constructing a series of 10 plasmids each with a 12 bp insertion in the gene encoding Vmw65. Plasmids were analysed for their ability to stimulate IE transcription in short term transfection assays, and the altered Vmw65 polypeptides were assayed for the ability to form an IE-specific protein-DNA complex (IEC) in vitro. A direct correlation was observed between stimulation of transcription and formation of IEC, strongly suggesting that IEC is an important intermediate in transcription activation. Plasmids were also tested for their ability to rescue the temperature-sensitive mutation in the HSV-2 assembly mutant ts2203, since marker rescue analysis indicated that this mutation maps within the gene encoding Vmw65. Five plasmids failed to rescue ts2203, thereby defining regions of Vmw65 required for virus assembly. The results show that distinct domains exist in Vmw65 for activation of transcription and assembly of virus.
单纯疱疹病毒1型（HSV - 1）的多肽Vmw65是病毒粒子的一种结构成分，并且还负责对立即早期（IE）转录进行反式诱导。通过构建一系列10个质粒来研究该多肽的功能结构域，每个质粒在编码Vmw65的基因中都有一个12 bp的插入片段。在短期转染实验中分析质粒刺激IE转录的能力，并在体外检测改变后的Vmw65多肽形成IE特异性蛋白质 - DNA复合物（IEC）的能力。观察到转录刺激与IEC形成之间存在直接相关性，强烈表明IEC是转录激活中的一个重要中间体。还测试了质粒拯救HSV - 2装配突变体ts2203中温度敏感突变的能力，因为标记拯救分析表明该突变位于编码Vmw65的基因内。五个质粒未能拯救ts2203，从而确定了病毒装配所需的Vmw65区域。结果表明，Vmw65中存在不同的结构域用于转录激活和病毒装配。
