ANALYSIS OF RETROVIRAL INTEGRASE COMPLEXES

将病毒DNA整合到宿主细胞的DNA中是 逆转录病毒的感染过程。这项提议的目的是澄清 病毒整合酶蛋白(IN)的寡聚体结构 对这一活动负责。IN蛋白已被证明是 DNA与LTR(长末端)整合的必要条件和充分条件 重复)末端在体外形成目标DNA。的性质和组成 将使用各种技术来阐明天然IN复合体 包括凝胶过滤、化学交联、天然凝胶 电泳法、圆二色谱和免疫沉淀法。单种氨基酸 将确定多聚体所需的序列，以及 经定点突变证实。功能分析也将是 用来检测这些突变对IN活性的影响。这个 逆转录病毒整合酶没有细胞对应物，因此代表 在感染过程中进行干预的合适目标。这些研究 将进一步加深我们对蛋白质结构如何与 逆转录病毒整合机制，并将有助于战略的设计 来控制逆转录病毒的复制。

The integration of viral DNA into the host cells' DNA is a required step in the retroviral infection process. The aim of this proposal is to elucidate the oligomeric structure of the viral integrase protein (IN), which is responsible for this activity. The IN protein has been shown to be necessary and sufficient for the integration of DNA with LTR (long terminal repeat) ends into a target DNA, in vitro. The nature and composition of native IN complexes will be elucidated using a variety of techniques including gel filtration, chemical cross-linking, native gel electrophoresis, CD spectra and immunoprecipitation. Individual amino acid sequences that are required for multimerization will be determined, and confirmed by site-directed mutagenesis. Functional assays will also be used to examine the effects of these mutations on IN activity. The retroviral integrase has no cellular counterpart, and therefore represents a suitable target for intervention in the infection process. These studies will further our understanding of how protein structure is related to the mechanism of retroviral integration, and will aid the design of strategies to control retroviral replication.