Everett R D
Institute of Virology, Glasgow, U.K.
J Mol Biol. 1988 Jul 5;202(1):87-96. doi: 10.1016/0022-2836(88)90521-9.
At the onset of infection by herpes simplex virus type 1 (HSV-1) in tissue culture, five immediate-early (IE) genes are expressed. At least four of the IE gene products have some role in the transcriptional regulation of later classes of viral genes. This study concerns the product of IE gene 1, Vmw110. Several laboratories have shown that a cloned copy of IE gene 1 can stimulate gene expression in trans in transfection experiments; this transactivation can occur either with Vmw110 by itself or in a strongly synergistic fashion with the product of IE gene 3, Vmw175. As an approach to study the mechanism of this effect, a large number of in-frame insertion mutations in the Vmw110 coding region have been constructed. The effect of the insertions on the ability of Vmw110 to activate transcription in the presence of Vmw175 has been described. This report concerns the effect of the insertions on the ability of Vmw110 to activate gene expression in the absence of Vmw175. In addition, a family of in-frame deletion mutations has been constructed and tested in transfection and immunofluorescence assays. The results show (1) that mutations in different domains of the Vmw110 polypeptide can have different effects on function in the presence or absence of Vmw175; (2) that mutations can affect the pattern of immunofluorescent intranuclear staining of Vmw110; and (3) that deletions in the region of a particular highly basic region of the polypeptide result in failure of Vmw110 to be transported into the nucleus.
在单纯疱疹病毒1型(HSV - 1)感染组织培养物开始时,会表达五个立即早期(IE)基因。至少有四个IE基因产物在后续几类病毒基因的转录调控中发挥某种作用。本研究关注的是IE基因1的产物Vmw110。几个实验室已经表明,在转染实验中,IE基因1的克隆拷贝可以反式刺激基因表达;这种反式激活既可以由Vmw110自身发生,也可以与IE基因3的产物Vmw175以强烈的协同方式发生。作为研究这种效应机制的一种方法,已经构建了大量Vmw110编码区的框内插入突变。已经描述了这些插入对Vmw110在存在Vmw175时激活转录能力的影响。本报告关注的是这些插入对Vmw110在不存在Vmw175时激活基因表达能力的影响。此外,还构建了一个框内缺失突变家族,并在转染和免疫荧光测定中进行了测试。结果表明:(1)在存在或不存在Vmw175的情况下,Vmw110多肽不同结构域的突变对其功能可能有不同影响;(2)突变可以影响Vmw110免疫荧光核内染色模式;(3)多肽特定高碱性区域的缺失导致Vmw110无法转运到细胞核中。