HIV Macromolecular Interactions and Impact on Viral Evolution of Drug Resistance

描述(由申请人提供)：我们已经成立了蜂巢中心，在原子水平上描述艾滋病毒生命周期中相互作用的大分子之间的结构和动态关系。我们将重点研究主要的艾滋病毒酶与它们的伴侣和效应物的相互作用，因为 它们包括病毒生命周期中的关键过程，作为现有的药物靶点，提供了丰富的结构、生物学和进化数据基础，将有助于实现我们的目标。我们将探索艾滋病毒的耐药性进化，以此为平台在原子水平上描述相互作用的大分子结构之间的动态关系。蜂巢中心由一组在艾滋病毒结晶学、病毒学、分子生物学、合成化学和计算生物学方面具有相当专业知识的研究人员组成。我们将研究病毒大分子相互作用和动力学的机制含义及其对耐药性演变的更广泛影响。该中心的目标是通过我们的研究回答以下基本问题：1.HIV多蛋白前体的结构如何指导组装、成熟和复制？2.什么新的HIV-宿主相互作用驱动DNA复制和整合？3.动力学如何影响病毒适合性，以及如何利用它进行治疗靶向？4.在HIV生命周期中，耐药突变的结构和动态后果是什么，以及如何？为了开展这些研究，我们将发展、改进和应用以下技术方法：1.HIV多蛋白和核酸复合体的高效表达、结晶和原子级结构测定。2.新结合位点的确定和表征及其对HIV蛋白复合体动态和进化特性的影响。3.在多时空尺度上表征HIV蛋白复合体相互作用、动力学和演化的计算方法。我们的方法对于承诺对病毒机制相互依赖的新的结构洞察具有重要意义，也对于新的药物设计方法和治疗策略的直接潜力具有重要意义。

DESCRIPTION (provided by applicant): We have formed the HIVE Center to characterize at the atomic level the structural and dynamic relationships between interacting macromolecules in the HIV life cycle. We will focus on interactions of the major HIV enzymes with their partners and effectors since they encompass key processes in the viral life cycle and as existing drug targets provide a rich base of structural, biological and evolutionary data that will serve to inform our goals. We will explore resistance evolution in HIV as an opportune platform upon which to characterize the dynamic relationships between interacting macromolecular structures at the atomic level. The HIVE Center comprises a group of investigators with considerable expertise in HIV crystallography, virology, molecular biology, synthetic chemistry and computational biology. We will study the mechanistic implications of viral macromolecular interactions and dynamics and its broader impacts of the evolution of drug resistance. The goals of this center are to answer the following fundamental questions with our studies: 1. How do structures of the HIV polyprotein precursors direct assembly, maturation, and replication? 2. What novel HIV-Host interactions drive DNA replication and integration? 3. How does dynamics impact viral fitness and how can it be exploited for therapeutic targeting? 4. What are the structural and dynamic consequences of resistance mutations in the HIV life cycle, and how? In order to carry out these studies we will develop, improve and apply the following Technical Methods for: 1. Efficient expression, crystallization, and atomic level structure determination of HIV polyprotein and nucleic acid complexes. 2. Determination and characterization of novel binding sites and their impact on the dynamic and evolutionary properties of HIV protein complexes. 3. Computational methods to characterize the interactions, dynamics and evolution of HIV protein complexes at multiple spatial and temporal scales. Our approach is significant both for the promise of new structural insights into the interdependence of viral mechanisms, but also for the direct potential for new drug design methodologies and therapeutic strategies.