Import of Adenovirus DNA into the nucleus

描述（由申请方提供）：腺病毒是基因组约为36 kb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。它们也是病毒性结膜炎的主要原因，包括流行性角膜结膜炎（EKC），这是一种可以威胁长期视觉功能的疾病，并且没有有效的治疗方法。此外，腺病毒由于其广泛的组织嗜性而被广泛研究作为人类基因治疗的载体。虽然腺病毒如何穿透细胞到达细胞质的重要见解已经获得，很少有人知道的腺病毒基因组的核输入的分子机制，这是病毒繁殖的关键。该建议旨在获得关于腺病毒DNA输入的详细分子见解。其目标是：1）研究腺病毒与核孔复合物对接的机制，重点分析腺病毒六邻体蛋白及其与特异性核孔蛋白的相互作用。2)将分析蛋白VII在腺病毒DNA通过核孔复合物运输中的作用，并研究蛋白VII可用作实现有效基因转移的非病毒方法的可能性。3)将分析胞质因子，包括hsc70及其辅因子，在孔复合物的病毒脱壳和DNA导入中的作用。综合考虑，这项工作将为了解致病性DNA病毒基因组的核输入提供一个有价值的模型。这项工作也可能促进人类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方法提供基础，这将有助于基因治疗和细胞功能研究。

DESCRIPTION (provided by applicant): 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.