MOLECULAR EVENTS AT THE HSV-1 ORIGIN OF DNA REPLICATION

HSV-1 DNA 复制起点的分子事件

基本信息

批准号：
2672546
负责人：
Paul E Boehmer
金额：
$ 19.51万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
1996
资助国家：
美国
起止时间：
1996-06-01 至 2000-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2672546
关键词：
Baculoviridae DNA binding protein DNA primase DNA replication origin Saccharomyces cerevisiae gene complementation gene mutation helicase herpes simplex virus 1 intermolecular interaction molecular cloning nucleic acid sequence protein purification protein sequence virus protein virus replication

项目摘要

The long term objectives of this research program are to understand the mechanisms by which the replication of herpes simplex virus type-1 (HSV-1) is initiated. Specific protein-protein interactions among the proteins engaged in this process are considered as targets for antiviral therapeutics. These studies should also provide insight into the mechanisms by which DNA replication is initiated from more complex eukaryotic chromosomal origins. HSV-1 is the prototype of the herpesviridae family of viruses. These viruses are known to cause a multi of disease in humans. HSV-1 in particular is extremely widespread in the population, and is associated with a number of symptoms that are a result of both primary and recurrent infections. The most prevalent conditions caused by infections with HSV-1 are oro-labial and genital skin lesions. However of much greater significance are encephalitis and blindness that are brought on by HSV-1 infections. Initiation of origin-specific DNA replication involves the targeted destabilization of particular chromosomal elements by a group of proteins that makes the DNA accessible to the DNA synthesis machinery, HSV-1 and single-stranded DNA binding protein (ICP8). The UL9 protein and ICP8 are known to form a tight complex that may function in origin recognition and unwinding. We hypothesize that the interaction between the UL9 protein and ICP8 is an essential part of the initiation process. Accordingly, we propose to further characterize the ICP8-UL9 protein complex. We will also determine the importance of this protein-protein interaction for origin- specific DNA replication. We further hypothesize that efficient activation of the origin of DNA replication involves the action of cellular proteins that assist the viral initiator proteins. Our approach is to identify such proteins by using biochemical and genetic complementation assays to detect specific protein-protein interactions between the viral initiator proteins and potential cellular factors. We propose to examine the importance of such cellular proteins by determining their role in activating the origin of DNA replication in vitro.

该研究计划的长期目标是了解 1 型单纯疱疹病毒 (HSV-1) 的复制机制已启动。蛋白质之间特定的蛋白质-蛋白质相互作用参与这一过程的人被视为抗病毒的目标疗法。这些研究还应该提供对 DNA复制从更复杂的机制开始真核染色体起源。 HSV-1 是疱疹病毒科病毒的原型。这些已知病毒会引起人类多种疾病。 HSV-1在特别是在人群中极为普遍，并且与具有许多由原发性和复发性原因引起的症状感染。由 HSV-1 感染引起的最常见病症是口唇和生殖器皮肤病变。然而更大的重要的是由 HSV-1 引起的脑炎和失明感染。起源特异性 DNA 复制的启动涉及目标一组蛋白质使特定染色体元件不稳定使 DNA 能够进入 DNA 合成机器、HSV-1 和单链 DNA 结合蛋白 (ICP8)。 UL9 蛋白和 ICP8 是已知形成紧密的复合物，可以在起源识别和放松。我们假设 UL9 蛋白与 ICP8 是启动过程的重要组成部分。据此，我们建议进一步表征 ICP8-UL9 蛋白质复合物。我们也会确定这种蛋白质-蛋白质相互作用对于起源的重要性- 特定的DNA复制。我们进一步假设有效激活 DNA复制起源涉及细胞蛋白质的作用协助病毒起始蛋白。我们的方法是识别这样的通过使用生化和遗传互补分析来检测蛋白质病毒起始蛋白之间的特异性蛋白质-蛋白质相互作用和潜在的细胞因素。我们建议研究以下问题的重要性：通过确定此类细胞蛋白在激活起源中的作用 DNA 体外复制。