Protein/Nucleic Acid Interactions Controlling Retroviral

抗病毒治疗的成功需要对参与HIV-1和HIV-2复制、组装和传染性的蛋白质有详细的了解。这些过程的中心是酶逆转录酶(RT)，十多年来它一直是抗病毒药物的主要靶点。由于RT介导的HIV RNA基因组复制是一个多步骤的过程，在分子水平上剖析和了解每个事件将产生扩大治疗靶点的结果。此外，目前的证据表明，RT可能不会单独作用于复制HIV基因组，而是招募更多的病毒蛋白。因此，更好地了解这些辅助蛋白和RT之间的“交流”可以为干预提供额外的途径。RT生物化学组的研究利用重组DNA技术准确地概括了HIV复制的步骤，并使用HIV-1、HIV-2和来自密切相关病毒的相应酶评估了它们对抗病毒药物的敏感性。此外，由于RT结构的改变导致耐药性的快速获得和抗病毒治疗的失败，了解这些细微变化的确切性质是很重要的。因此，除了生化分析外，该实验室目前还应用生物偶联和质谱学方法，努力获得这种高度通用的酶的野生型和突变型的高分辨率结构信息。

The success of antiviral therapies will require a detailed knowledge of proteins involved in the replication, assembly, and infectivity of HIV-1 and HIV-2. Central to these processes is the enzyme reverse transcriptase (RT), which has been a leading target for antiviral drugs for over a decade. Since RT-mediated replication of the HIV RNA genome is a multistep process, dissecting and understanding each event at the molecular level will have the consequence of expanding the repertoire of therapeutic targets. Moreover, current evidence suggests RT may not work alone in replicating the HIV genome, but rather recruits additional viral proteins. A better understanding of the "communication" between these accessory proteins and RT can therefore offer additional avenues for intervention. Research in the RT Biochemistry Section exploits recombinant DNA technology to accurately recapitulate steps in HIV replication and evaluate their sensitivity to antiviral agents, using the enzymes of HIV-1, HIV-2, and their counterparts from closely related viruses. Furthermore, since alterations in RT structure result in the rapid acquisition of drug resistance and failure of antiviral therapies, it is important to understand the exact nature of these subtle alterations. Thus, in addition to a biochemical analysis, the laboratory currently applies bioconjugate and mass spectrometric approaches in an effort to obtain high-resolution structural information on both wild-type and mutant variants of this highly versatile enzyme.