Import of Adenovirus DNA into the nucleus

将腺病毒 DNA 导入细胞核

• 批准号: 7160516
• 负责人: Larry R. Gerace
• 金额: $ 35.59万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7160516
• 关键词: Adenovirus hexon capsid protein; Adenoviruses; Binding; Biochemical; Cell Nucleus; Cells; Clinical; Complex; Condition; Cultured Cells; Cytoplasm; DNA; DNA Viruses; Development; Docking; Epidemic Keratoconjunctivitis; Eye Infections; Gene Transduction Agent; Gene Transfer; Genome; Goals; Human; Human Adenoviruses; Methods; Modeling; Molecular; Nuclear Import; Nuclear Pore Complex; Nuclear Pore Complex Proteins; Numbers; Proteins; Reproduction; Respiratory Tract Infections; Role; Viral Conjunctivitis; Virus; Vision; Work; base; cofactor; gastrointestinal; gene therapy; insight; respiratory; tissue tropism; vector

Adenoviruses are non-enveloped DNA viruses with an -36 kb genome. In humans, adenoviruses cause a significant number of gastrointestinal and respiratory infections. They also are a major cause of viral conjunctivitis including epidemic keratoconjunctivitis (EKC), a condition that can threaten long-term visual function and for which there is no effective treatment. In addition, adenoviruses are being intensively investigated as vectors for human gene therapy because of their broad tissue tropism. Although significant insight has been obtained on how adenovirus penetrates the cell to reach the cytoplasm, little is known about the molecular mechanism of nuclear import of the adenovirus genome, which is critical for virus reproduction. This proposal is directed at obtaining detailed molecular insight on adenovirus DNA import. The aims are: 1) The mechanism for docking of adenovirus to the nuclear pore complex will be investigated, focusing on an analysis of the adenovirus hexon protein and its interaction with specific nucleoporins. 2) The role of protein VII in the transport of adenovirus DNA through the nuclear pore complex will be analyzed, and the possibility that protein VII can be used as a nonviral method for achieving efficient gene transfer will be investigated. 3) The role of cytosolic factors, including hsc70 and its cofactors, in virus uncoating at the pore complex and in DNA import will be analyzed. Considered together, this work will provide a valuable model for understanding the nuclear import of the genomes of pathogenic DNA viruses. The work also could potentiate the development of new therapies for EKC in humans. Finally, it could provide the basis for developing efficient means for nonviral gene transfer, which would be useful for gene therapy and functional studies of cells.

腺病毒是具有 -36 kb 基因组的无包膜 DNA 病毒。在人类中，腺病毒 引起大量胃肠道和呼吸道感染。他们也是一大 病毒性结膜炎的病因，包括流行性角结膜炎 (EKC)，这种疾病可以 威胁长期视觉功能，并且没有有效的治疗方法。此外， 腺病毒作为人类基因治疗的载体正在得到深入研究，因为它们具有以下特点： 广泛的组织趋向性。尽管对于腺病毒如何渗透已获得重要见解 细胞到达细胞质的过程中，对于细胞核输入的分子机制知之甚少。 腺病毒基因组，对于病毒繁殖至关重要。该提案针对的是 获得有关腺病毒 DNA 导入的详细分子见解。目标是： 1) 机制 将研究腺病毒与核孔复合物的对接，重点是分析 腺病毒六邻体蛋白及其与特定核孔蛋白的相互作用。 2）蛋白质的作用 VII 腺病毒 DNA 通过核孔复合物的运输将被分析，并且 蛋白质VII可用作实现有效基因转移的非病毒方法的可能性将 被调查。 3) 胞质因子，包括hsc70及其辅因子，在病毒脱壳中的作用 将分析孔复合物和 DNA 输入中的情况。综合考虑，这项工作将提供 一个了解致病性 DNA 病毒基因组核输入的有价值的模型。 这项工作还可以促进人类 EKC 新疗法的开发。最后，它 可以为开发有效的非病毒基因转移手段提供基础，这将是 可用于基因治疗和细胞功能研究。