Structure and Evolution of APOBEC3-Vif Interactions

APOBEC3-Vif 相互作用的结构和演化

基本信息

批准号：
10229568
负责人：
John D Gross
金额：
$ 46.6万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-08-27 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10229568
关键词：
26S proteasome 3-Dimensional Allosteric Site Binding Proteins Biochemistry Biological Assay CUL5 gene Cells Cercocebus Cercopithecidae Collaborations Complementary DNA Complex Core-Binding Factor Cryoelectron Microscopy Crystallization Cytidine Deaminase Data Enzymes Evolution Fab Immunoglobulins Family Family Study Family member Genes Genetic Genetic Transcription Genome HIV HIV-1 Hominidae In Vitro Individual Innate Immune System Length Lentivirus Maps Methods Modeling Molecular Monkeys Mutagenesis Mutation Negative Staining Pan Genus Polynucleotides Polyubiquitin Primates Proteins RNA Binding Race Recording of previous events Retroelements Retroviridae Reverse Transcription SIV Structure Surface System Testing Time Ubiquitin Ubiquitination Viral Viral Genome Viral Packaging Virion Virus Virus Replication X-Ray Crystallography arm c new cofactor cross-species transmission in vitro Assay insight multicatalytic endopeptidase complex novel particle pathogen pathogenic virus receptor reconstitution reconstruction restraint ubiquitin-protein ligase vif Gene Products vpr Genes

项目摘要

Restriction factors are a branch of the innate immune system that potently inhibit viral replication. A number of viral pathogens encode accessory proteins that can antagonize restriction factors, which allows viral dissemination in host. Over evolutionary time, restriction factors evolve to escape viral antagonists, limiting the host range of viruses. In turn, viruses can adapt to these changes and cross species into new hosts by rapid evolution of accessory proteins. The primate APOBEC3 cytidine deaminases are arguably the most extensively studied family of restriction factors. They block the spread of retroviruses and retroelements by hypermutating their genomes and have the potential to inhibit the AIDS virus, HIV-1. However, the HIV-1 accessory protein Vif potently suppresses APOBEC3 enzymes by targeting them for degradation by the host ubiquitin-proteasome system. Vif is conserved in all existing lentiviruses, including those of primate origin such as SIV. Adaptation in Vif has allowed cross-species transmission of SIV from monkeys to chimpanzees and underlies the ancient origin of HIV-1. Accordingly, the APOBEC3-Vif interaction is an archetypical case of a restriction factor and viral antagonist. In principle, host escape by mutation in APOBEC3 and viral adaptation in Vif could occur through changes in protein binding interfaces, insertion of short-linear interaction motifs by gene loss and overprinting or mutation of allosteric sites. However, there are no co-structures of Vif bound to any APOBEC3 family member to document these phenomena, so the detailed mechanisms are unclear. In this project, we will overlay sequencing and viral infectivity data accumulated at critical points during the evolutionary history of HIV-1 with structures of corresponding Vif-A3 complexes. Cutting edge methods, such as Fab-assisted single-particle cryo-EM, will be used to resolve these structures. The functional significance of the observed interactions will be tested by mutagenesis in conjunction with viral infectivity assays in cells and Vif ubiquitination activity assays in vitro. This project will provide a paradigmatic example of the biochemistry, structure, and molecular mechanisms of molecular arms races that are general phenomena of host-pathogen interactions.

限制因素是有效抑制病毒复制的先天免疫系统的一个分支。许多病毒病原体编码可以拮抗限制因子的辅助蛋白，这允许病毒在主持人中传播。在进化的时间内，限制因素演变为逃避病毒拮抗剂，限制了宿主病毒范围。反过来，病毒可以通过快速的附件蛋白的演变。灵长类动物apobec3 cytidine脱氨酶可以说是最大的广泛研究了限制因素家族。它们通过过度渗透其基因组，并具有抑制AIDS病毒HIV-1的潜力。但是，HIV-1 辅助蛋白VIF通过靶向宿主降解来有效抑制APOBEC3酶泛素 - 蛋白酶体系统。 VIF在所有现有的慢病毒中均保存，包括灵长类动物起源的病毒作为SIV。 VIF中的适应已允许SIV从猴子传播到黑猩猩和是HIV-1的古代起源。因此，apobec3-vif相互作用是一个典型的情况限制因子和病毒拮抗剂。原则上，宿主在ApoBec3中突变逃脱，并在 VIF可能通过蛋白质结合界面的变化，插入短线相互作用图案的变化而发生基因丧失和变构位点的过度打印或突变。但是，没有VIF的共同结构与任何APOBEC3家庭成员都记录了这些现象，因此详细的机制尚不清楚。在这个项目，我们将覆盖测序和病毒感染数据在临界点积累 HIV-1具有相应VIF-A3复合物结构的进化史。尖端方法，这样作为Fab辅助的单粒子冷冻EM，将用于解决这些结构。功能意义的观察到的相互作用将通过诱变与细胞中的病毒感染分析结合进行测试 vif泛素化活性在体外进行。该项目将提供生物化学的范式示例，分子臂种族的结构和分子机制是宿主病原体的一般现象互动。