ADENO-ASSOCIATED VIRUS IN VITRO INTEGRATION AND REPLICATION

腺相关病毒的体外整合和复制

• 批准号: 5203534
• 负责人: R KOTIN
• 金额: --
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/5203534
• 关键词: DNA directed DNA polymerase; DNA replication; adeno associated virus group; gene duplication; gene rearrangement; genetic recombination; intermolecular interaction; microorganism culture; nucleic acid repetitive sequence; provirus; virus DNA; virus genetics; virus infection mechanism; virus protein; virus replication

Adeno-associated virus (AAV) DNA integrates preferentially into a specific locus on human chromosome 19 by non-homologous recombination. The interactions of the non-structural viral protein, Rep, with chromosome 19-derived DNA and AAV internal terminal repeats (ITR) suggests involvement of the viral protein in targeted integration. We propose that targeted integration results from limited DNA replication initiated by binding of Rep to a consensus binding sequencing present in both the integration locus and the viral ITR. We postulate that a dynamic mechanism involving both limited DNA synthesis of the cellular sequences and protein- protein interactions between subunits of Rep protein and a cellular DNA polymerase complex generates the viral-cellular recombination junctions. The generalized and extended Rep functions we have determined are consistent with this model. Analysis of genomic DNA of latently infected cell lines are supportive as well in that duplications and rearrangements of cellular sequences are present. Furthermore, every AAV provirus characterized to date was present asymmetrically distributed with respect to the Rep-binding element.

腺相关病毒（AAV）DNA优先整合到一个 通过非同源重组在人19号染色体上的特异性基因座。 非结构性病毒蛋白Rep与 19号染色体衍生的DNA和腺相关病毒内部末端重复序列（ITR） 表明病毒蛋白参与靶向整合。 我们 我认为靶向整合是由有限的DNA复制引起的， 通过Rep与存在的共有结合序列的结合来启动 在整合位点和病毒ITR中。 我们假设a 动力学机制涉及细胞的有限DNA合成， Rep亚基之间的序列和蛋白质-蛋白质相互作用 蛋白质和细胞DNA聚合酶复合物产生 病毒-细胞重组连接。 广义和广义 我们确定的代表函数与该模型一致。 潜伏感染细胞系的基因组DNA分析是支持性的 细胞序列的复制和重排 都在场 此外，迄今为止表征的每种AAV前病毒都是 相对于Rep结合呈现不对称分布 元素