Host cell regulation of Adenovirus gene expression

腺病毒基因表达的宿主细胞调控

• 批准号: 6954500
• 负责人: Eileen K BRIDGE
• 金额: $ 21.3万
• 依托单位: MIAMI UNIVERSITY OXFORD
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6954500
• 关键词: Adenoviridae; DNA virus; cell line; gene expression; genetic regulation; host organism interaction; immediate early protein; immunofluorescence technique; interferons; mutant; protein localization; protein structure function; transcription factor; tumor suppressor proteins; virus RNA; virus genetics; virus infection mechanism; virus protein; virus replication; western blottings

DESCRIPTION (provided by applicant): Adenovirus (Ad) is a linear double-stranded DNA virus. Successful lytic infection by Ad requires a productive interaction with its host cell, creating an environment that is conducive to efficient viral gene expression, replication, and assembly of new progeny virus. Ad regulatory proteins derived from early region 4 (E4), E4-11K and E4-34K, are important in cell transformation, viral replication, and late gene expression; they are also able to interact with host double-strand break repair (DSBR) proteins and affect their localization and stability. E4 mutants have their genomes "repaired" in infected cells to form concatemers of several genome lengths. Here, we propose to investigate the role of host DSBR processes in the replication and late gene expression defects displayed by E4 mutants lacking E4-11K and E4-34K. The experiments will investigate the relationship between the function of E4 proteins in inactivating DSBR, and their role in promoting viral replication and gene expression. We will study the significance of genome concatenation for viral late gene expression in E4 mutant infections. We will study the role of individual DSBR proteins and genome concatenation in the onset of E4 mutant DNA replication. The ability of Ad genomes to induce a damage response in the presence and absence of viral DNA replication will be assessed. Finally, we will study the role of nuclear domain 10 (ND10) disruptions for viral replication. ND10 contains DSBR proteins that are targeted by E4 regulatory proteins. Our goals are to investigate possible links between the ability of E4 proteins to inactivate host DSBR, and their function in promoting viral replication and gene expression during lytic infections. The project has long term significance for understanding how viral genes affect host cell activities, and how they may ultimately affect cell growth and transformation as they reprogram the machinery of the host to promote a successful lytic viral infection.

描述（申请人提供）： 腺病毒（Ad）是一种线性双链DNA病毒。 Ad 成功的裂解感染需要与其宿主细胞进行有效的相互作用，创造一个有利于病毒基因高效表达、复制和新子代病毒组装的环境。源自早期区域 4 (E4)、E4-11K 和 E4-34K 的 Ad 调节蛋白在细胞转化、病毒复制和晚期基因表达中非常重要；它们还能够与宿主双链断裂修复（DSBR）蛋白相互作用并影响其定位和稳定性。 E4突变体的基因组在受感染的细胞中被“修复”，形成多个基因组长度的串联体。在这里，我们建议研究宿主 DSBR 过程在缺乏 E4-11K 和 E4-34K 的 E4 突变体复制和晚期基因表达缺陷中的作用。这些实验将研究E4蛋白在灭活DSBR中的功能与它们在促进病毒复制和基因表达中的作用之间的关系。我们将研究基因组串联对于 E4 突变体感染中病毒晚期基因表达的重要性。我们将研究单个 DSBR 蛋白和基因组串联在 E4 突变体 DNA 复制开始时的作用。将评估 Ad 基因组在存在和不存在病毒 DNA 复制的情况下诱导损伤反应的能力。最后，我们将研究核结构域 10 (ND10) 破坏对病毒复制的作用。 ND10 包含 E4 调节蛋白靶向的 DSBR 蛋白。我们的目标是研究 E4 蛋白灭活宿主 DSBR 的能力与其在裂解感染期间促进病毒复制和基因表达的功能之间可能的联系。该项目对于了解病毒基因如何影响宿主细胞活动，以及它们如何重新编程宿主机制以促进成功的裂解性病毒感染，最终如何影响细胞生长和转化具有长期意义。