Enzymatic Mechanism of Herpes Virus DNA Replication

本研究的目的是了解（i）疱疹病毒DNA复制启动的机制，（ii） 疱疹病毒DNA复制过程中发生倒位的机制，以及（iii） 影响疱疹病毒感染的神经细胞中病毒潜伏期的发展。我们预计， 研究将为我们提供一个深入了解的复制，重组和潜伏期的一个重要类别， 人类病原体调查将按以下方式沿着进行。 A.多聚体来源结合蛋白（UL 9蛋白）与Oris结合的刺激分析， HSV-1 DNA复制起点，通过细胞hTid-1 DnaJ分子伴侣。 B。神经F盒蛋白（NFB 42）在HSV- 1潜伏期中的作用 1.结合NFB 42后UL 9蛋白的泛素化和降解。 2.研究UL 9蛋白与NFB 42相互作用的磷酸化状态。 C.核酸内切酶G在HSV- 1基因组序列倒位起始中的作用。 1.核酸内切酶G切割一个序列的研究。 2. HSV- 1感染对核酸内切酶G水平的影响。 3. HSV-1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.