Enzymatic Mechanism of Herpes Virus DNA Replication

这项研究的目的是了解(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的发展。我们预计这些 研究将为我们提供对重要类别的复制、重组和延迟的洞察 人类病原体。调查工作将按照以下思路进行。 A.多聚体来源结合蛋白(UL9蛋白)与Oris、And结合刺激的分析 HSV-1 DNA复制的起源，由细胞内的hTid-1 DNAJ伴侣。 B.神经F盒蛋白(NFB42)在HSV-1潜伏期中的作用 1.结合NFB42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。

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 Oris, 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 NFB42. 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.