Structure and Function of HIV-1 Reverse Transcriptase

HIV-1逆转录酶的结构和功能

基本信息

批准号：
7733047
负责人：
stephen h hughes
金额：
$ 94.71万
依托单位：
DIVISION OF BASIC SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

项目摘要

HIV-1 is the causative agent of AIDS. Three viral enzymes - reverse transcriptase (RT), integrase (IN), and protease (PR) - have essential roles in the replication of HIV-1. We are engaged in a long-term effort to study HIV-1 RT, with the expectation that this information will be useful in the development of more effective anti-RT drugs. Our strategy has involved the analysis of both wild-type and mutant HIV-1 RTs, including drug-resistant mutants. Some of this purified RT has been used by our long-term collaborator, Dr. Eddy Arnold (Rutgers University), for structural studies. We have used purified HIV-1 RT to study the biochemical properties of RT mutants, including drug-resistant mutants. There are two clinically important classes of inhibitors of HIV-1 RT: nucleoside analogs (NRTIs) and nonnucleoside inhibitors (NNRTIs). Both are used to treat HIV-1 infections; however, there are serious problems with drug toxicity and with the development of resistance. A major focus of our work on HIV-1 RT is the mechanism(s) of RT inhibitor resistance. We will continue to investigate the mechanisms that underlie drug resistance; however, we have begun to direct a part of our efforts to the development of novel inhibitors that will be effective against the known drug-resistant RTs. We have a collaboration with a skilled nucleoside chemist, Dr. Victor Marquez (Laboratory of Medicinal Chemistry, NCI); the goal of this collaboration is to develop nucleoside analogs that are relatively resistant to excision by NRTI-resistant HIV-1 RTs that are excision proficient. A part of our collaboration with Dr. Arnold is intended to develop more effective NNRTIs. All of the experiments designed to understand NRTI and NNRTI resistance and to develop more effective inhibitors involve a combined structural/biochemical approach; what we have learned from RT structure has been an invaluable guide for planning the biochemical studies, while the results and hypotheses generated in the biochemical experiments have inspired new rounds of structural experiments. Drug resistance is only one aspect of the behavior of RT. We also want to understand how RT carries out reverse transcription in an infected cell, and to correlate the wealth of structural and biochemical data on HIV-1 RT with the actual process of reverse transcription. These experiments are part of Project Z01 BC 010482 (Retroviral Replication and Vector Design). In some cases, it will not be possible to explain the in vivo data with the available structural and biochemical results. In such cases, we will do additional biochemical and structural experiments to complement (and better understand) the in vivo results. [Corresponds to Hughes Project 1 in the April 2007 site visit report of the HIV Drug Resistance Program]

HIV-1是艾滋病的病因。三种病毒酶 - 逆转录酶（RT），整合酶（IN）和蛋白酶（PR） - 在HIV -1的复制中具有重要作用。我们从事长期研究HIV-1 RT的努力，期望此信息可用于开发更有效的抗RT药物。我们的策略涉及对野生型和突变体HIV-1 RT的分析，包括耐药突变体。我们的长期合作者Eddy Arnold（Rutgers University）使用了一些纯化的RT进行结构研究。我们已经使用纯化的HIV-1 RT来研究RT突变体的生化特性，包括抗药性突变体。 HIV-1 RT的抑制剂有两种临床上重要的类别：核苷类似物（NRTIS）和非核苷抑制剂（NNRTIS）。两者都用于治疗HIV-1感染；但是，药物毒性和抗药性发展存在严重问题。我们在HIV-1 RT上的主要重点是RT抑制剂耐药性的机制。我们将继续研究抗药性基础的机制；但是，我们已经开始将一部分努力引向开发新型抑制剂，这些抑制剂将有效地抵抗已知的耐药性RT。我们与熟练的核苷化学家Victor Marquez博士（NCI药物实验室）合作；这种合作的目的是开发核苷类似物，这些核苷类似物对熟练的切除的NRTI-NRTI耐药HIV-1 RT相对抗性。我们与Arnold博士合作的一部分旨在开发更有效的NNRTIS。所有旨在了解NRTI和NNRTI耐药性并开发更有效抑制剂的实验均涉及一种结构性/生化方法。我们从RT结构中学到的是计划生化研究的宝贵指南，而生化实验中产生的结果和假设启发了新的结构实验。耐药性只是RT行为的一个方面。我们还想了解RT如何在受感染的细胞中进行逆转录，并将HIV-1 RT上的结构和生化数据的财富与逆转录的实际过程相关联。这些实验是项目Z01 BC 010482（逆转录病毒复制和矢量设计）的一部分。在某些情况下，不可能用可用的结构和生化结果来解释体内数据。在这种情况下，我们将进行其他生化和结构实验，以补充体内结果（并更好地理解）。 [对应于休斯项目1在2007年4月的现场访问艾滋病毒抗药性计划的报告中]