Molecular Biology of APOBECS and Vif Interactions

APOBECS 和 Vif 相互作用的分子生物学

• 批准号: 8433370
• 负责人: Reuben S Harris
• 金额: $ 49.14万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8433370
• 关键词: AIDS/HIV problem; APRIN gene; Amino Acids; Anti-Retroviral Agents; Binding; Binding Sites; Biochemical; Biochemistry; Biological; Biological Assay; C-terminal; CD4 Positive T Lymphocytes; Catalytic Domain; Cell Count; Cells; Charge; Collaborations; Complementary DNA; Complex; Conflict (Psychology); DNA; DNA Binding; Data Set; Deaminase; Development; Dimerization; EMSA; Electrostatics; Genetic; HIV; HIV therapy; HIV-1; Human; Kinetics; Knowledge; Laboratories; Life; Maps; Mediating; Modeling; Molecular; Molecular Biology; Molecular Probes; Molecular Virology; Mutagenesis; Nature; Pathogenesis; Pathway interactions; Patients; Pharmaceutical Preparations; Property; Proteins; Reporting; Single-Stranded DNA; Structure; Surface; Testing; Therapeutic; Viral; Work; acrosome stabilizing factor; base; design; dimer; forging; in vivo; inhibitor/antagonist; monomer; mutant; novel; pathogen; prevent; programs; protein protein interaction; single molecule; small molecule; vif Gene Products

The discovery of the anti-viral APOBECS proteins is one of the most therapeutically promising breakthroughs in HIV/AIDS molecular virology in recent years. Humans have seven APOBECS proteins and at least two, APOBECSG and APOBECSF, are capable of inhibiting the replication of Vif-deficient HIV. However, HIV pathogenesis is due at least in part to the fact that the viral Vif protein counteracts these APOBECS proteins and triggers their degradation. The molecular approaches described in this proposal are an integral part of a larger program project to provide comprehensive knowledge of the structural, biophysical, biochemical and molecular features of these APOBECS proteins and their relation to HIV-1. First, we will test the hypothesis that APOBECSG dimerization occurs through multiple direct protein-protein interactions, and we will determine the relevance of these interactions to HIV restriction. Second, we will distinguish between two models for how APOBECSG binds single-strand DNA substrates analogous to HIV cDNA. Third, we will test the hypothesis that HIV Vif recognizes a common structural motif that is present in APOBECSF, APOBECSG, and other APOBECS proteins. We will use structural information from our program collaborations to guide the construction of APOBECS and Vif mutants. These studies will be aided in part by a panel of novel APOBECSG inhibitory small molecules that will be used as molecular probes to dissect these critical steps of the AP0BEC3G/F-mediated HIV-1 restriction mechanism. These studies will advance our fundamental understanding of APOBECSG and APOBECSF and facilitate the development of novel HIV/AIDS therapeutics that work by modulating the APOBECS-Vif pathway.

近年来，抗病毒Apobecs蛋白的发现是艾滋病毒/艾滋病分子病毒学中最有希望的突破之一。人类具有七种Apobecs蛋白质，至少两种APOBECSG和APOBECSF能够抑制VIF缺陷HIV的复制。但是，HIV发病机理至少部分是由于病毒VIF蛋白抵消这些Apobecs蛋白并触发其降解的事实。该提案中描述的分子方法是一个较大程序项目的组成部分，旨在全面了解这些APOBEC蛋白的结构，生物物理，生化和分子特征及其与HIV-1的关系。首先，我们将检验以下假设：apobecsg二聚化是通过多种直接蛋白质 - 蛋白质相互作用发生的，我们将确定这些相互作用与HIV限制的相关性。其次，我们将区分两种模型，即APOBECSG如何结合类似于HIV cDNA的单链DNA底物。第三，我们将检验以下假设：HIV VIF识别APOBECSF，APOBECSG和其他Apobecs蛋白中存在的常见结构基序。我们将使用计划合作中的结构信息来指导APOBEC和VIF突变体的构建。这些研究将由一组新型的APOBECSG抑制性小分子进行帮助，这些小分子将用作分子探针，以剖析AP0BEC3G/F介导的HIV-1限制机制的这些关键步骤。这些研究将提高我们对APOBECSG和APOBECSF的基本理解，并促进通过调节Apobecs-VIF途径来起作用的新型HIV/AIDS治疗剂的发展。