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， 因此可以在其他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。 对现有抗艾滋病毒疗法的耐药性继续上升。要继续成功地抑制艾滋病毒在受感染者体内的复制，就需要开发出能有效对抗耐药病毒株的新抑制剂。靶向病毒复制新区域的药物最有可能对这些病毒有效。拟议的工作重点是在早期HIV-1感染细胞过程中形成的大型病毒DNA（vDNA）复合物-逆转录和整合前复合物（RTC和PIC）。这些复合物促进病毒DNA的合成及其转运到细胞核中。开发新抑制剂的第一步是确定这些复合物的组成。与RTC和PIC相关的病毒蛋白与其大小不一致，表明它们含有额外的细胞组分。我们的长期目标是通过确定RTC和PIC如何组装和介导vDNA在分裂和非分裂细胞中的核转运来帮助开发新的HIV-1抑制剂。拟议研究的目的是确定和表征在HIV-1复制中发挥重要作用的HIV-1 vDNA复合物的新细胞组分。我们已经开发了新的方法来纯化这些复合物，我们已经成功地检测到病毒成分的质谱。使用这些方法，我们将确定新的宿主蛋白，是HIV-1复制的关键。这些研究的成功完成将有助于了解艾滋病毒复制的早期事件，并发现抗病毒治疗的新靶点。

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. Drug resistance to the existing pool of anti-HIV therapies continues to rise. Continued success in the repression of HIV replication in infected individuals will require the development of new inhibitors that are effective against drug-resistant strains of virus. Drugs that target novel areas of virus replication have the greatest probability to be effective against such viruses. The proposed work focuses on the large viral DNA (vDNA) complexes formed during early HIV-1 infection of cells- the reverse transcription and preintegration complexes (RTCs and PICs). These complexes facilitate synthesis of viral DNA and its transport into the nucleus of cells. The first step toward the development of new inhibitors is to determine the composition of these complexes. The viral proteins associated with RTCs and PICs are not consistent with their size, suggesting they contain additional cellular components. Our long-term goal is to aid in the development of new HIV-1 inhibitors by determining how RTCs and PICs assemble and mediate nuclear transport of vDNA in dividing and non-dividing cells. The objective of the proposed studies is to identify and characterize novel cellular components of HIV-1 vDNA complexes that play an essential role in HIV-1 replication. We have developed novel methods to purify these complexes and we have successfully detected viral components by mass spectrometry. Using these methods we will identify novel host proteins that are critical for HIV-1 replication. Successful completion of these studies will contribute to the understanding of early events of HIV replication and discover new targets for antiviral therapies.