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Molecular basis of herpes simplex vius pathogeniciby

单纯疱疹病毒致病的分子基础

基本信息

批准号：
05454199
负责人：
NISHIYAMA Yukihiro
金额：
$ 3.65万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

项目摘要

1. The protein kinase encoded by the US3 gene of herpes simplex virus type 2 (HSV-2) was purified more than 1000-fold from the post-ribosomal supernatant of infected Vero cells, and the final preparation contained one major protein of apparent molecular weight 66K,which was phosphorylated in the autophosphorylation reaction. The HSV-2 protein kinase was relatively resistant to high concentrations of salt, and when the substrate specificity was investigated using synthetic oligopeptides, the peptides containing arginyl residues on the amino-terminal side of the target residue were found to be the best substrates for the US3 protein kinase.2. When permeabilized cells were labelled with [gamma-^<32>P] ATP under the optimum conditions for the US3 protein kinase, the most striking difference between wild-type and US3-inactivated virus-infected cells was observed in the phosphorylation of proteins ranging in Mr values from 14K to 21K.The results indicate that a tegment phosphoprotein encoded by the US9 gene may be a target of the enzyme.Similar studies demonstrate that the alkaline nuclease encoded by UL12 was also a major target of the US3 protein kinase, and suggest that phosphorylation by the protein kinase may be involved in the stability/activity of the alkaline nuclease in murine macrophages.3. We have studied the pathogenic and latency/reactivation potential of a herpes simplex virus type 1 (HSV-1) variant (N38) which has a deletion of four genes, US9,10,11 and 12, in the short unique region of the HSV-1 genome. N38 was a pathogenic as the parental wild-type virus following intracerebral infection of mice and replicated with wild-type kinetics in the brain. There was no significant difference between two viruses in the frequency of reactivation or in reactivation time. The results indicate that these four genes are dispensable for its neurovirulence and latency in mice.

1.从感染Vero细胞的核糖体上清液中纯化了单纯疱疹病毒2型(HSV-2)US3基因编码的蛋白激酶，纯化倍数达1000多倍，最终产物中含有一个表观分子量为66K的主要蛋白，该蛋白在自磷酸化反应中被磷酸化。结论：1.HSV-2蛋白激酶对高浓度盐有较强的耐受性，当使用合成的寡肽研究底物特异性时，目标残基的氨基末端含有精氨酸残基的多肽被发现是US3蛋白激酶的最佳底物。当通透性细胞在US3蛋白激酶的最佳条件下用[Gamma-^&^&lt；32&gt；P]ATP标记时，野生型和US3灭活病毒感染细胞之间最显著的差异是蛋白质的磷酸化，范围从14K到21K。结果表明，US9基因编码的被盖磷酸蛋白可能是酶的靶标。类似的研究表明，UL12编码的碱性核酸酶也是US3蛋白激酶的主要靶标，提示蛋白激酶的磷酸化可能参与了小鼠巨噬细胞碱性核酸酶的稳定/活性。我们研究了单纯疱疹病毒1型(HSV-1)变异株(N38)的致病性和潜伏期/再激活潜能，该变异株在HSV-1基因组的短独特区中缺失了US9、10、11和12四个基因。N38是一种致病病毒，与小鼠脑内感染的亲本野生型病毒一样，在脑内以野生型动力学方式复制。两种病毒在重新激活的频率或重新激活时间上没有显著差异。结果表明，这四个基因对小鼠的神经毒力和潜伏期是必不可少的。