STRUCTURE FUNCTION RELATIONSHIP IN HIV RT

逆转录酶是许多致癌过程中不可或缺的一部分 病毒，包括来自人类白血病和艾滋病病毒的病毒， 对于病毒复制来说是必不可少的。这方面的长期目标 建议是理解和定义生物化学， 逆转录酶的酶学和结构性质 因此，逆转录的分子机制可以 被澄清了。因此，这项研究的主要目标是 鉴定和确定逆转录酶的结构域 它们参与底物dNTPs与模板的结合。 以及那些负责反向表达的引物。 转录酶相关核糖核酸酶H。结果在预料之中。 阐明酶促合成DNA的基本机制 由逆转录酶催化，并可能提供对 独特的特性/结构，可能有助于设计 潜在的治疗剂。 我们分析了抗逆转录酶活性的机制。 一些抑制剂，并已确定了一些试剂 似乎与特定部位的逆转录酶反应 举止。此外，我们还对共价协议进行了标准化 将底物、模板底物和特定试剂连接到 酶蛋白。这些试剂在机构中很有用。 RT的位点特异性共价修饰，进而将 允许携带多肽的标签、鉴定和分离 这些网站。通过这种方式，我们希望确定域 负责MuLV和HIV的聚合酶和核糖核酸酶H活性 逆转录酶。这方面的知识将对 了解多种抗病毒药物的确切机制(包括 抗艾滋病)具有假定的抗RT活性的药物。

Reverse transcriptase is an integral part of many oncogenic viruses, including those from human leukemia and AIDS virus, which is essential for viral replication. Long range goals of this proposal are to understand and define the biochemical, enzymological and structural properties of reverse transcriptase so that molecular mechanisms underlying reverse transcription could be clarified. The major objective, therefore, of this research is to identify and define structural domains of reverse transcriptase which are involved in the binding of substrate dNTPs and template- primer as well as those responsible for the expression of reverse- transcriptase associated Ribonuclease H. The results are expected to clarify the basic mechanisms of enzymatic synthesis of DNA catalyzed by reverse transcriptase and may provide an insight into unique properties/structures which may help in designing the potential therapeutic agents. We have analyzed mechanism of anti-reverse transcriptase activity of a number of inhibitors and have identified some reagents which appear to react with reverse transcriptase in a site-specific manner. In addition, we have standardized protocols for covalently linking substrates, template-primers and specific reagents to the enzyme protein. These reagents are quite useful in establishment of site-specific covalent modification of RT which in turn will permit labeling, identification and isolation of peptides carrying these sites. In this manner, we hope to identify domains responsible for the polymerase and RNAse H activity of MuLV and HIV reverse transcriptase. This knowledge will be very useful in understanding of precise mechanisms of many anti-viral (including anti-AIDS) drugs with presumptive anti-RT activity.