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Studies on the properties and functions of accessory gene products of herpes simplex virus

单纯疱疹病毒副基因产物的特性和功能研究

基本信息

批准号：
09470083
负责人：
NISHIYAMA Yukihiro
金额：
$ 7.55万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09470083/
关键词：
herpes simplex virus accessory gene function of viral proteins protein kinase mechanism of viral replication 非必須遺伝子 US3 アポトーシス

项目摘要

We have constructed prokaryotic and eukaryotic expression vectors of herpes simplex virus type 2 (HSV-2) genes, generated rabbit polyclonal antibodies against fusion proteins expressed in Escherichia coli, and then identified a number of HSV-2 gene products including UL3, UL4, UL14, UL16, UL17, UL31, UL51, UL55, and US2. All of them were produced at the late phase of infection : properties of some of them are summarized as follows.・The UL4 gene product was identified as a 27-kDa protein in infected cells. Indirect immunofluorescence studies localized the UL14 protein within the nucleus as discrete punctate forms at late times postinfection, but the protein was limited to the cytoplasm when expressed alone, indicating that an interaction with one or more other virus-induced proteins was responsible for the nuclear localization during infection.・The UL16 gene product was identified as a 41-kDa protein in infected cells, and produced in a manner highly dependent on viral DNA synthesis. The UL16 protein was distributed in both the nuclei and the cytoplasm, and was associated with intracellular C capsids. Our results suggest that the UL16 protein plays a role in capsid maturation including DNA packaging/cleavage.We have also studied the biological role of the US3 gene product in a murine model of HSV infection, and found that the US3 protein kinase plays an important role in protecting HSV-2-infected cells from apoptotic death in vivo.

我们构建了单纯疱疹病毒2型（HSV-2）基因的原核和真核表达载体，制备了兔抗大肠杆菌表达融合蛋白的多克隆抗体，并鉴定了一些HSV-2基因产物，包括UL 3，UL 4，UL 14，UL 16，UL 17，UL 31，UL 51，UL 55和US 2。它们都是在感染后期产生的：其中一些的特性总结如下。·UL 4基因产物在感染细胞中被鉴定为27 kDa蛋白质。间接免疫荧光研究将UL 14蛋白定位在细胞核内，在感染后的后期以离散的点状形式存在，但当单独表达时，该蛋白仅限于细胞质，这表明与一种或多种其他病毒诱导蛋白的相互作用是感染期间细胞核定位的原因。UL 16基因产物在感染细胞中被鉴定为41-kDa蛋白，并且以高度依赖于病毒DNA合成的方式产生。UL 16蛋白分布于细胞核和细胞质中，并与细胞内的C衣壳相关。我们的研究结果表明，UL 16蛋白在衣壳成熟，包括DNA包装/切割中发挥作用。我们还研究了US 3基因产物在HSV感染的小鼠模型中的生物学作用，并发现US 3蛋白激酶在体内保护HSV-2感染的细胞免于凋亡中发挥重要作用。