Protein-Protein Interactions of HIV-1 Integrase

超出所提供的空间。该项目的长期目标是阐明宿主与病毒的动态相互作用，特别是整合酶（IN）和INI1/hSNF5，并利用这些知识设计抗艾滋病治疗药物。INI1是染色质重塑SWI/SNF复合物的核心成分，也是一种肿瘤抑制因子。INI1/hSNF5被特异性地整合到HIV-1病毒粒子中，INI1/hSNF5显性的截断突变体负性地抑制HIV-1复制。INI1/hSNF5携带一个掩蔽的核输出信号，介导依赖于hCRM1/ exportl的核输出。我们发现INI1/hSNF5也存在于另一种新的复合体中，不同于SWI/SNF复合体，其成分被并入HIV-1病毒粒子中。我们假设，在组装和颗粒生产过程中，INI1/hSNF5及其新的核复合体被信号输出到细胞质，停靠在Gag-Pol的IN部分，并组装成HIV-1病毒粒子。在靶细胞中，INI1复合体可能促进原病毒的整合和/或调节转录。在这个建议中，在特定的目标1中，我们将使用RNA干扰和转显性突变体来确定INI 1和SAP 18是否需要在产生细胞或靶细胞中进行HIV-1复制的早期和晚期事件。在特定目的II中，我们将描述inil相关蛋白（如SAP18）及其与HIV-1病毒粒子结合的机制。我们将纯化新的INI1-SAP18多蛋白复合物，并确定其对IN的依赖性。在具体目标III中，我们将进行生化和遗传分析，以确定INI1及其相关蛋白如SAP18对HIV-1复制的功能意义。我们将测试纯化的INI 1及其复合物对体外整合的影响，分离和表征in、INI 1和SAP 18的相互作用缺陷突变体及其分子内和分子间、等位基因特异性和非等位基因特异性的回复性。在具体目的IV中，我们将确定INI1/hSNF5的核输出特性是否对HIV-1病毒粒子的掺入以及早期和晚期事件的跨显性抑制很重要。我们的研究将全面阐述inil相关多蛋白复合物募集HIV-1的机制和意义，并阐明IN与其宿主蛋白的高阶结构/功能关系。了解宿主-病毒相互作用的活力可能为设计有效的抗HIV-1抗病毒药物提供新的靶点和见解。网站性能 ======================================== 节结束 ===========================================

EXCEED THE SPACE PROVIDED. The long-term objective of this project is aimed at elucidating the dynamic host-virus interactions, specifically focusing on integrase (IN) and INI1/hSNF5, and to utilize this knowledge for the design of drugs for anti-AIDS therapy. INI1 is a core component of the chromatin remodeling SWI/SNF complex and a tumor suppressor. INI1/hSNF5 is specifically incorporated into the HIV-1 virions and a truncation mutant of INI1/hSNF5 dominant negatively inhibits HIV-1 replication. INI1/hSNF5 harbors a masked nuclear export signal that mediates the hCRM1/exportinl dependent nuclear export. We have found that INI1/hSNF5 is also present in another novel complex, distinct from the SWI/SNF complex, the component of which is incorporated into HIV-1 virions. We hypothesize that during assembly and particle production, INI1/hSNF5, along with its novel nuclear complex, is signaled to get exported to the cytoplasm, docks itself onto IN portion of Gag-Pol, and gets assembled into the HIV-1 virions. In the target cells, INI1 complex may facilitate the integration and/or regulate transcription of the provirus. In this proposal, in the specific aim I, using RNA interference and trasdominant mutants we will determine if INI 1 and SAP 18 are required for early and late events of HIV-1 replication in producer cells or target cells. In the specific aim II, we will characterize INIl-assocated proteins (such as SAP18) and mechanism of their incorporation into HIV-1 virions. We will purify the novel INI1-SAP18 multi-protein complex and determine the dependence on IN. In the specific aim III, we will carry out biochemical and genetic analysis to determine the functional significance of INI1 and its associated proteins such as SAP18 to HIV-1 replication. We will test the effect of purified INI 1 and complexes on in vitro integration, isolate and characterize interaction defective mutants of IN, INI 1 and SAP 18 and their intra-molecular and inter-molecular, allele specific and non-allele specific revertants. In the specific aim IV, we will determine if nuclear export property of INI1/hSNF5 is important for incorporation into HIV-1 virions and for transdominant inhibition of early and late events. Our proposal will provide a comprehensive account of mechanism and significance of recruitment of INIl-associated multi-protein complex to HIV-1 and elucidate the higher order structure/function relationship of IN with its host proteins. Understanding the vitality of host-virus interactions is likely to provide novel targets and insights for designing effective antivirals against HIV-1. PERFORMANCE SITE ========================================Section End===========================================