APOBEC3 STRUCTURAL STUDIES

APOBEC3 结构研究

基本信息

批准号：
9332852
负责人：
Hideki Aihara
金额：
$ 7.96万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
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/9332852
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 Human Immune In Vitro Individual Knowledge Lead Light Macromolecular Complexes Mediating Methods Modeling Molecular Molecular Virology Mutagenesis Mutation Natural Immunity Patients Polyubiquitination Proteins Publishing Reaction Recruitment Activity Regimen Resolution Reverse Transcription Roentgen Rays SIV Series Single-Stranded DNA Structure Subfamily lentivirinae Surface Testing Therapeutic Ubiquitination Uracil Viral Virion Virus Virus Replication Work antiretroviral therapy atomic interactions base combinatorial crosslink curative treatments cytosine analog genome integrity inhibitor/antagonist innovation insight novel pandemic disease particle pathogen protein complex public health relevance research study structural biology ubiquitin ligase ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): 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.

描述（由适用提供）：抗病毒APOBEC3酶的发现被认为是HIV/AIDS分子病毒学中最有希望的突破之一。几种APOBEC3s可以通过纳入组装病毒颗粒，物理干扰逆转录的进展，并将病毒cDNA细胞固醇脱离到尿嘧啶中，从而限制HIV复制。后一种抗病毒活性是APOBEC3介导的限制的定义标志，解释了在患者衍生的病毒序列中经常观察到的基因组链G-TO-A突变。然而，这些潜在的抗病毒活性由HIV病毒体感染因子（VIF）抵消，该因子与CBF-beta异二聚体，以形成E3泛素连接酶复合物，从而降解Apobec3酶。在这里，我们将解决该领域的两个持久问题。首先，我们这些结构将为结合APOBEC3结合的VIF/CBF-BETA泛素连接酶配合物提供见解。这些结构将提供有关APOBEC3-VIF结合机制，构象变化以及这些宿主病原体复合物的整体组织的见解。其次，我们将阐明APOBEC3/单链DNA复合物的X射线结构。这些结构将提供有关酶催化，DNA底物识别以及DNA结合和催化活性释放之间的偶联机制的见解。我们预计，来自APOBEC3/VIF/CBF-BETA和APOBEC3/单链DNA大分子复合物的结构信息在从长远来看至关重要，因为该领域继续朝着这些相互作用进行吸毒并通过天然的先天免疫组织实现潜在的HIV-1限制。