APOBEC3 STRUCTURAL STUDIES

APOBEC3 结构研究

• 批准号: 9726634
• 负责人: Hideki Aihara
• 金额: $ 3.52万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-15 至 2020-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9726634
• 关键词: AIDS/HIV problem; Active Sites; Address; Affinity; Amino Acids; Anti-Retroviral Agents; Antiviral Agents; Binding; Biochemical; Biological; Biological Assay; CUL5 gene; Catalysis; Chemicals; Complementary DNA; Complex; Core-Binding Factor; Coupling; Crystallization; Cytosine; DNA; DNA Binding; Data; Deamination; Dinucleoside Phosphates; Disulfides; Drug Interactions; Drug resistance; Engineering; Enzymes; Event; Evolution; Future; Genetic Transcription; Genomics; HIV; HIV Infections; HIV-1; Heterodimerization; Human; Human immunodeficiency virus test; Immune; In Vitro; Individual; Knowledge; Lead; Light; Macromolecular Complexes; Mediating; Methods; Modeling; Molecular; Molecular Conformation; Molecular Virology; Mutagenesis; Mutation; Mutation Analysis; Natural Immunity; Patients; Polyubiquitination; Proteins; Publishing; Reaction; Regimen; Resolution; Reverse Transcription; Roentgen Rays; SIV; Series; Single-Stranded DNA; Structure; Subfamily lentivirinae; Surface; Testing; Therapeutic; Uracil; Viral; Virion; Virus; Virus Replication; Work; antiretroviral therapy; base; combinatorial; crosslink; curative treatments; cytosine analog; experimental study; genome integrity; inhibitor/antagonist; innovation; insight; novel; pandemic disease; particle; pathogen; protein complex; recruit; structural biology; ubiquitin ligase; ubiquitin-protein ligase; virology

APOBEC3 STRUCTURAL STUDIES ABSTRACT The discovery of the antiviral APOBEC3 enzymes is regarded as one of the most therapeutically promising breakthroughs in HIV/AIDS molecular virology. Several APOBEC3s have the potential to restrict HIV replication by incorporating into assembling viral particles, physically interfering with the progression of reverse transcription, and deaminating viral cDNA cytosines to uracils. The latter antiviral activity is the defining hallmark of APOBEC3-mediated restriction, explaining the genomic strand G-to-A mutations that are frequently observed in patient-derived viral sequences. However, these potent antiviral activities are counteracted by the HIV virion infectivity factor (Vif), which heterodimerizes with CBF-beta in order to form an E3 ubiquitin ligase complex that degrades APOBEC3 enzymes. Here, we will address two persisting problems in this field. First, we will determine x-ray structures of APOBEC3-bound Vif/CBF-beta ubiquitin ligase complexes. These structures will provide insights into the APOBEC3-Vif binding mechanism, conformational changes, and the overall organization of these host-pathogen complexes. Second, we will elucidate X-ray structures of APOBEC3/single-stranded DNA complexes. These structures will provide insights into the mechanisms of enzymatic catalysis, DNA substrate recognition, and coupling between DNA-binding and the release of catalytic activity. We anticipate that structural information from both APOBEC3/Vif/CBF-beta and APOBEC3/single-stranded DNA macromolecular complexes will be important in the longer-term as the field continues to move toward drugging these interactions and enabling potent HIV-1 restriction through natural innate immunity.

APOBEC 3结构研究 摘要 抗病毒APOBEC 3酶的发现被认为是最有治疗前景的酶之一。 艾滋病毒/艾滋病分子病毒学的突破。几种APOBEC 3具有限制HIV的潜力 通过掺入组装的病毒颗粒中，物理干扰逆转录病毒的进展， 转录，并将病毒cDNA胞嘧啶脱氨为尿嘧啶。后者的抗病毒活性是定义 APOBEC 3介导的限制的标志，解释了基因组链G到A突变， 在患者来源的病毒序列中观察到。然而，这些有效的抗病毒活性被抗病毒药物所抵消。 HIV病毒体感染因子（Vif），与CBF-β异源二聚化以形成E3泛素连接酶 复合物，降解APOBEC 3酶。在这里，我们将解决这一领域的两个长期存在的问题。第一、 我们将确定APOBEC 3结合的Vif/CBF-β泛素连接酶复合物的X射线结构。这些 结构将提供深入了解APOBEC 3-Vif结合机制，构象变化， 这些宿主-病原体复合体的整体组织。其次，我们将阐明的X射线结构的 APOBEC 3/单链DNA复合物。这些结构将提供深入了解的机制， 酶催化，DNA底物识别，以及DNA结合和释放之间的偶联， 催化活性。我们预期来自APOBEC 3/Vif/CBF-beta和 APOBEC 3/单链DNA大分子复合物将是重要的，在长期的领域， 继续朝着药物这些相互作用，使有效的HIV-1限制，通过自然的， 先天免疫