CHARACTERIZATION OF HIV-1 PREINTEGRATION COMPLEX ASSEMBLY AND NUCLEAR TRANSPORT

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 耐药株的演变对人类免疫缺陷病毒1型（HIV-1）感染的有效长期治疗构成了重大挑战。针对耐药菌株的一种对策是用新疗法进行治疗。HIV复制的几个领域是开发新疗法的潜在目标。我们研究的目的是阐明HIV前整合复合物（PIC）的组装和转运，以帮助开发新的抗逆转录病毒疗法。PIC是在HIV-1早期感染细胞期间形成的大的病毒DNA复合物，其在逆转录后将病毒DNA靶向细胞核。由于生产和纯化足够量的PIC的挑战，对其组成、组装和运输机制的理解有限。因此，没有开发中的PIC抑制剂。我们研究的目的是识别和表征HIV-1 PIC的新细胞成分。我们已经采取了两个雄心勃勃的战略，以确定PIC相关的细胞蛋白。首先是通过速度梯度离心部分纯化的PIC的靶向蛋白质组学分析。第二，是通过HIV-1蛋白的特异性生物素化纯化的PIC亲和力的蛋白质组学分析。使用这两种方法，我们已经确定了几个候选人的HIV依赖因子（HDFs）。将评估候选HDF与HIV组分的相互作用及其在HIV复制中的作用。这些研究的成功完成将促进对艾滋病毒复制早期事件的了解，并发现开发抗病毒疗法的新靶点。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The evolution of drug resistant strains poses a significant challenge for effective long-term treatment of human immunodeficiency virus type 1 (HIV-1) infection. One counter-measure against resistant strains is treatment with novel therapeutics. Several areas of HIV replication are potential targets for the development of new therapies. The goal of our research is to elucidate the assembly and transport of the HIV preintegration complex (PIC) to aid in the development of novel antiretroviral therapeutics. PICs are large viral DNA complexes formed during early HIV-1 infection of cells that target the viral DNA into the nuclei of cells after reverse transcription. Due to challenges in producing and purifying sufficient quantities of PICs there is limited understanding of their composition, assembly, and mechanism of the transport. Consequently, there are no inhibitors of PICs in development. The objective of our studies is to identify and characterize novel cellular components of HIV-1 PICs. We have undertaken two ambitious strategies to identify PIC-associated cellular proteins. First is a targeted proteomic analysis of PICs partially purified by velocity gradient centrifugation. Second, is a proteomic analysis of PICs affinity purified by specific biotinylation of HIV-1 proteins. Using both methods we have identified several candidate HIV dependency factors (HDFs). Candidate HDFs will be assessed for their interaction with HIV components and their role in HIV replication. Successful completion of these studies will advance the understanding of early events of HIV replication and discover new targets for the development of antiviral therapies.