Cellular DNAJ Proteins and SV40 Infection

细胞 DNAJ 蛋白和 SV40 感染

• 批准号: 7726053
• 负责人: Daniel C. Dimaio
• 金额: $ 24.09万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7726053
• 关键词: BK Virus; Binding; Biochemical; Biological; Cancer Patient; Capsid; Cell surface; Cells; Collaborations; Disease; Escape Mutant; Event; Family; Family member; Funding; Gene Expression; Gene Family; Genes; Genetic; Genetic Screening; Grant; Head; Human; Human Herpesvirus 4; Human Virus; Human papillomavirus 18; Individual; Infection; Large T Antigen; Lead; Molecular; Molecular Chaperones; Monkeys; Oncogenic Viruses; Papillomavirus; Play; Polyomavirus; Polyomavirus Infections; Process; Property; Proteins; Relative (related person); Repression; Role; Series; Signal Transduction; Simian virus 40; Technology; Testing; Transmembrane Transport; Tumor Virus Infections; Viral; Virus; Virus Diseases; base; cellular targeting; immunosuppressed; insight; knock-down; member; metaplastic cell transformation; mutant; novel; novel strategies; prevent; reconstitution; research study; response; senescence; small hairpin RNA; trafficking

This new project headed by Dr. DiMaio is based on a discovery made in project 2 during the current funding period. The polyomaviruses, BK virus andJC virus, cause serious diseases in immunosuppressed individuals including cancer patients and, like their close relative SV40, are putative human tumor viruses. With the support of this grant, we discovered that the cellular co-chaperones DNAJ-B12 and DNAJ-B14 are required for efficient infection by these three viruses. When expression of these genes is repressed by shRNAs, there is a substantial reduction in expression of the major early protein, large T antigen. Virus binding to the cell surface appears unimpaired, suggesting that the DNAJ-B12/14 sensitive step(s) is in some aspect of virus entry, intracellular trafficking, or uncoating. We will conduct a series of biochemical, cell biological and genetic studies to elucidate the mechanistic role played by DNAJ-B12/14 in SV40 infection. We will determine how far infection proceeds in cells lacking DNAJ-B12/14 function, and determine what step in the virus entry/trafficking/uncoating process is blocked. We will conduct mutational and biochemical analysis of DNAJ-B12/14 to determine its mode of action at the molecular level. Viral escape mutants that allow infection despite DNAJ-B12/14 repression will be isolated and characterized. Finally, we will use shRNA technology to determine whether other members of the DNAJ gene family are required for infection by the polyomaviruses and other viruses, including Epstein-Barr virus in collaboration with Dr. Miller. These experiments will provide new insights into the process of tumor virus infection and characterize the role of new putative anti-viral targets.

这个由DiMaio博士领导的新项目是基于项目2在当前 资助期。多瘤病毒，BK病毒和JC病毒，在免疫抑制的情况下导致严重的疾病 包括癌症患者在内的个人和他们的近亲SV40一样，都是假定的人类肿瘤病毒。 在这项资助的支持下，我们发现细胞共伴侣DNAJ-B12和DNAJ-B14是 有效感染这三种病毒所必需的。当这些基因的表达被抑制时 ShRNAs，主要的早期蛋白，大T抗原的表达大幅减少。病毒 与细胞表面的结合未见损伤，提示DNAJ-B12/14敏感步骤(S)在某些情况下 病毒进入、细胞内传播或脱膜的方面。我们将进行一系列的生化、细胞 生物学和遗传学研究，阐明DNAJ-B12/14在SV40感染中的作用机制。 我们将确定在缺乏DNAJ-B12/14功能的细胞中感染进行了多远，并确定 病毒进入/传播/去涂层过程中的步骤被阻止。我们将进行突变和生化 分析DNAJ-B12/14，在分子水平上确定其作用模式。病毒逃逸突变体 尽管DNAJ-B12/14抑制，但允许感染将被分离和鉴定。最后，我们将使用 ShRNA技术用于确定感染是否需要DNAJ基因家族的其他成员 由多瘤病毒和其他病毒，包括与米勒博士合作的爱泼斯坦-巴尔病毒。这些 实验将为肿瘤病毒感染的过程提供新的见解，并表征 新的假定的抗病毒靶点。