HPV helper functions for AAV Replication

用于 AAV 复制的 HPV 辅助函数

• 批准号: 6872478
• 负责人: PAUL L HERMONAT
• 金额: $ 23.29万
• 依托单位: UNIV OF ARKANSAS FOR MED SCIS
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-15 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6872478
• 关键词: DNA replication; adeno associated virus group; cervix neoplasms; gene therapy; genetic transcription; helper virus; human papillomavirus; human tissue; transfection /expression vector; viral carcinogenesis; virion; virus genetics; virus protein; virus replication

DESCRIPTION (provided by applicant): This proposal will analyze the helper functions of HPV for Adeno-Associated Virus (AAV) replication. The parvovirus AAV has a complex life cycle, as a helper-dependent virus, as an autonomous virus in skin, or as an integrated provirus. It is known that adenoviruses, herpesviruses, and human papillomaviruses (HPV) can serve as helpers for AAV. While the adenovirus and herpesvirus helper genes have been studied, HPV helper genes are largely unstudied. We now find that of the HPV early genes E1 has the highest ability to help AAV replication. Our goal in this proposal is to characterize the extent and mechanism of E1 helper function for AAV. The knowledge we gain on E1 helper function for AAV will have dual utility: 1) AAV and HPV interact in skin and their interaction modulates the risk of cervical cancer. Understanding how HPV helps AAV replication, and the mechanism of action, will provide new insight into the cervical carcinogenesis process. 2) Second, rAAV is gaining increasing popularity as a human gene therapy vector. Understanding HPV helper functions for AAV should provide alternative strategies for rAAV production. To analyze HPV helper function for AAV we will: Aim #1: Determine if HPV E1 regulates AAV DNA replication, transcription, and virion production. Aim #2: Investigate the mechanism of E1 helper function by mapping the "AAV helper domain" and "Rep78- interaction domain" within the HPV E1 protein. E1 mutants will be used to map the protein domains. Knowing the helper and Rep78 interaction domains within E1 protein will be the first step in deciphering El's mechanism of helper action. Aim #3: Investigate El's mechanism of action by observing cooperation/ complementation of E1 helper function with the known adenovirus helper genes.

描述（由申请人提供）：该提案将分析 HPV 对于腺相关病毒（AAV）复制的辅助功能。 细小病毒 AAV 具有复杂的生命周期，作为辅助依赖性病毒、作为皮肤中的自主病毒或作为整合的原病毒。众所周知，腺病毒、疱疹病毒和人乳头瘤病毒 (HPV) 可以作为 AAV 的辅助病毒。虽然腺病毒和疱疹病毒辅助基因已被研究，但 HPV 辅助基因很大程度上尚未被研究。我们现在发现HPV早期基因E1具有最高的帮助AAV复制的能力。我们在此提案中的目标是描述 AAV 的 E1 辅助功能的范围和机制。我们获得的关于 AAV E1 辅助功能的知识将具有双重用途：1) AAV 和 HPV 在皮肤中相互作用，并且它们的相互作用调节宫颈癌的风险。了解 HPV 如何帮助 AAV 复制及其作用机制，将为了解宫颈癌发生过程提供新的见解。 2) 其次，rAAV 作为人类基因治疗载体越来越受欢迎。了解 AAV 的 HPV 辅助功能应该为 rAAV 生产提供替代策略。 为了分析 AAV 的 HPV 辅助功能，我们将： 目标#1：确定 HPV E1 是否调节 AAV DNA 复制、转录和病毒粒子产生。目标#2：通过绘制 HPV E1 蛋白内的“AAV 辅助结构域”和“Rep78 相互作用结构域”来研究 E1 辅助功能的机制。 E1 突变体将用于绘制蛋白质结构域图。了解 E1 蛋白内辅助蛋白和 Rep78 相互作用结构域将是破译 El 辅助作用机制的第一步。目标#3：通过观察 E1 辅助功能与已知腺病毒辅助基因的合作/互补来研究 El 的作用机制。