APOBEC3 STRUCTURAL STUDIES

APOBEC3 结构研究

基本信息

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
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/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

项目摘要

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.

APOBEC3 结构研究抽象的抗病毒 APOBEC3 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 APOBEC3 具有限制 HIV 的潜力通过掺入组装病毒颗粒进行复制，物理干扰反向进程转录，以及将病毒 cDNA 胞嘧啶脱氨基为尿嘧啶。后者的抗病毒活性是决定性的 APOBEC3 介导的限制的标志，解释了经常发生的基因组链 G 至 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会被 HIV 病毒粒子感染因子 (Vif)，与 CBF-β 异二聚化以形成 E3 泛素连接酶降解 APOBEC3 酶的复合物。在这里，我们将解决该领域两个长期存在的问题。第一的，我们将确定 APOBEC3 结合的 Vif/CBF-β 泛素连接酶复合物的 X 射线结构。这些结构将提供对 APOBEC3-Vif 结合机制、构象变化和这些宿主-病原体复合物的整体组织。其次，我们将阐明其X射线结构。 APOBEC3/单链 DNA 复合物。这些结构将提供对机制的见解酶催化、DNA 底物识别以及 DNA 结合和释放之间的耦合催化活性。我们预计 APOBEC3/Vif/CBF-beta 和从长远来看，APOBEC3/单链DNA大分子复合物将在该领域发挥重要作用继续努力对这些相互作用进行药物治疗，并通过自然方式实现有效的 HIV-1 限制先天免疫。