Identification of novel co-factors for HIV Tat and Rev as therapeutic targets

描述(申请人提供)：这个项目的目标是确定HIV-1感染的新治疗靶点，重点是病毒TAT中涉及的细胞辅助因素 蛋白质或Rev蛋白的功能。虽然目前还不知道有多少细胞因子参与了TAT和REV的功能，但最近的研究和数据挖掘表明，大约有50种细胞蛋白可能参与了这些过程。对TAT最了解的细胞辅助因子是一种被称为P-TEFb的蛋白激酶复合体。P-TEFb由细胞周期蛋白T1(Cyclin T1，CCNT1)作为调节亚基，CDK9作为催化亚基组成，Tat直接与CCNT1结合。对REV最了解的细胞辅助因子称为XPO1(CRM1)；REV直接与XPO1结合。细胞中几乎所有的过程都是由被认为是最小功能单位的多蛋白复合体完成的。因此，CCNT1/CDK9和XPO1都可能是几个高阶的多蛋白复合体，这些复合体可能参与了TAT和REV的功能。我们挖掘了最近发表的描述人类核复合体的数据集-高通量免疫共沉淀/质谱仪分析定义的一组多蛋白复合体。我们已经发现了一些含有CCN1/CDK9或XPO1的新型细胞复合体。我们将测试这样的假设，即这些复合体中的几个参与TAT或REV功能。在我们的初步研究中，我们已经获得了对这一假说的强有力的实验支持。重要的是，我们的初步结果还表明，我们将识别对HIV-LTR的TAT激活进行负面调节的细胞因素。这些负面作用因素在清除服用抗病毒药物患者的潜在HIV-1储存库方面具有治疗潜力。拟议工作的完成将揭示一些潜在的治疗目标，特别是一些可能有助于清除艾滋病毒感染者体内潜伏病毒的战略。 公共卫生相关性：我们将确定调节HIV-1复制的细胞蛋白。其中一些蛋白质可能是治疗感染的有用靶点。我们的研究可能会改善患者的治疗。

DESCRIPTION (provided by applicant): The goal of this project is to identify novel therapeutic targets for HIV-1 infection, focusing on cellular co- factors involved in the viral Tat protein or Rev protein function. Although it is not known how many cellular factors are involved in Tat and Rev function, recent studies and data mining have revealed that on the order of 50 cellular proteins are likely involved in these processes. The best understood cellular co-factor for Tat is a protein kinase complex termed P-TEFb. P-TEFb is composed of Cyclin T1 (CCNT1) as a regulatory subunit and CDK9 as the catalytic subunit; Tat binds directly to CCNT1. The best understood cellular co-factor for Rev is termed XPO1 (CRM1); Rev binds directly to XPO1. Nearly all processes in cells are carried out by multiprotein complexes which are thought to be the minimal functional unit. It is therefore likely that both CCNT1/CDK9 and XPO1 exist is several higher order multiprotein complexes and these complexes may be involved in Tat and Rev function. We have mined a recently published dataset that described the human nuclear complexome - the set of multiprotein complexes defined by a high-throughput co-immunoprecipitation/mass spec analysis. We have identified a number of novel cellular complexes that contain CCN1/CDK9 or XPO1. We will test the hypothesis that several of these complexes are involved in either Tat or Rev function. In our preliminary studies, we have obtained strong experimental support for this hypothesis. Importantly, our initial results also indicate that we will identify cellular factors that negatively regulate Tat activation of the HIV- LTR. Such negative acting factors have therapeutic potential in strategies to purge the latent HIV-1 reservoir in patients taking anti-viral drugs. Completion of the proposed work will reveal a number of potential therapeutic targets, especially some that may be useful for strategies to purge the reservoir of latent viruses in HIV-infected individuals. PUBLIC HEALTH RELEVANCE: We will identify cellular proteins that regulate HIV-1 replication. Some of these proteins might be useful as therapeutic targets to treat infection. Our research may improve patient treatment.