Enzymatic Mechanism of Herpes Virus DNA Replication

描述(由申请人提供)： 本研究的目的是了解(I)疱疹病毒DNA复制的启动机制，(Ii)在疱疹病毒DNA复制过程中发生倒置的机制，以及(Iii)影响疱疹病毒感染神经细胞中病毒潜伏期发展的因素。我们预计，这些研究将为我们提供对一类重要的人类病原体的复制、重组和潜伏时间的洞察。调查工作将按照以下思路进行。 A.分析细胞hTid-1DNAJ伴侣对单纯疱疹病毒1DNA复制来源之一的Ori-S结合多聚体来源结合蛋白的刺激作用。 B.神经F盒蛋白(NFB42)在HSV-1潜伏期中的作用 1.结合NFB4后UL9蛋白的泛素化和降解 2.研究UL9蛋白与NFB42相互作用的磷酸化状态。 C.内切酶G在HSV-1基因组序列倒置启动中的作用。 1.核酸内切酶G切割序列的研究 2.单纯疱疹病毒1型感染对核酸内切酶G的影响 3.单纯疱疹病毒1型感染前后核酸内切酶G的细胞定位 4.核酸内切酶G与其他蛋白质的相互作用。 5.核酸内切酶G的抑制对α-序列介导的重组的影响。

DESCRIPTION (provided by applicant): The aim of this research is to understand (i) the mechanism of initiation of herpes virus DNA replication, (ii) the mechanism of inversion that occurs during herpes virus DNA replication and (iii) the factors that influence the development of viral latency in herpes virus-infected neuronal cells. We anticipate that these studies will provide us with an insight into the replication, recombination and latency of a significant class of human pathogens. The investigation will be organized along the following lines. A. Analysis of the stimulation of binding of multimeric origin binding protein (UL9 protein) to Ori-s, an origin of HSV-1 DNA replication, by the cellular hTid-1 DnaJ chaperone. B. Role of neural F box protein (NFB42) in HSV- 1 latency. 1. Ubiquitination and degradation of UL9 protein upon binding NFB4 2. Investigation of state of phosphorylation of UL9 protein on its interaction with NFB42. C. Role of endonuclease G in the initiation of sequence inversion in the HSV- 1 genome. 1. Studies of a sequence cleavage by endonuclease G. 2. Effect of HSV- 1 infection on levels of endonuclease G. 3. Cellular localization of endonuclease G pre- and post HSV-1 infection 4. Interaction of endonuclease G with other proteins. 5. Effect of suppression of endonuclease G on a-sequence-mediated recombination.