Structural studies of biomolecules related to cancer and HIV-AIDS

与癌症和艾滋病相关的生物分子的结构研究

基本信息

批准号：
10703074
负责人：
Hiroshi Matsuo
金额：
$ 69.5万
依托单位：
DIVISION OF BASIC SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
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/10703074
关键词：
APOCEC3G gene Acquired Immunodeficiency Syndrome Affinity Amino Acids Binding Catalytic Domain Cells Complex Conflict (Psychology)Cryoelectron Microscopy Crystallization Cytosine deaminase DNA DNA Sequence Alteration Defense Mechanisms Electron Microscopy Epitopes Evolution Goals HIV HIV Genome HIV-1 HIV/AIDS Human Infection Magnetism Malignant Neoplasms Modification Mutate Mutation Nuclear Nucleotides Pathway interactions Prognosis Proteins Publishing RNA Reaction Research Research Project Grants Role Signal Transduction Single-Stranded DNA Source Spectrum Analysis Stable Isotope Labeling Structure Techniques Testing Ubiquitin Virion X-Ray Crystallography Zebularine analog base cancer therapy cancer type inhibitor multicatalytic endopeptidase complex novel therapeutics pathogen protein structure response therapy resistant tumor tumor growth tumor progression ubiquitin-protein ligase

APOCEC3G gene Acquired Immunodeficiency Syndrome Affinity Amino Acids Binding Catalytic Domain Cells Complex Conflict (Psychology)Cryoelectron Microscopy Crystallization Cytosine deaminase DNA DNA Sequence Alteration Defense Mechanisms Electron Microscopy Epitopes Evolution Goals HIV HIV Genome HIV-1 HIV/AIDS Human Infection Magnetism Malignant Neoplasms Modification Mutate Mutation Nuclear Nucleotides Pathway interactions Prognosis Proteins Publishing RNA Reaction Research Research Project Grants Role Signal Transduction Single-Stranded DNA Source Spectrum Analysis Stable Isotope Labeling Structure Techniques Testing Ubiquitin Virion X-Ray Crystallography Zebularine analog base cancer therapy cancer type inhibitor multicatalytic endopeptidase complex novel therapeutics pathogen protein structure response therapy resistant tumor tumor growth tumor progression ubiquitin-protein ligase

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项目摘要

APOBEC3G (A3G) and APOBEC3H (A3H) are single-stranded DNA cytosine deaminase that can restrict HIV-1 infection by mutating HIV-1 genome. HIV-1 developed a counter defense mechanism by which virion infectivity factor (Vif) leads the degradation of A3G/A3H through ubiquitin-proteasome pathway. Our ultimate goal is to generate small compounds which inhibit the degradation of A3G/A3H. Previously, we had determined NMR and crystal structures of domains of A3 proteins, including the VIf-binding domain and the catalytic domain. In addition, we had determined co-crystal structures of the A3's catalytic domain-ssDNA complex. During 2021-2022, we have developed a technique to selectively stable-isotope label Vif in complex with A3G, and assigned NMR signals of Vif to identify amino acid residues in the interfaces with A3G (published in JMR, 2022). We have made progress in determining the structure of the A3H-Vif E3 ubiquitin ligase complex by using cryoEM, which will provide epitopes to be targeted by small compounds which inhibit formation of the complex. Dysregulation of APOBEC3A (A3A) and APOBEC3B (A3B) proteins contributes a major endogenous source of DNA mutations traced in approximately 75% of cancer types and 50% of all cancers analyzed. A3A/A3B proteins can cause DNA mutations either alone or as the response to cancer therapies which can drive evolution of cancers, and A3A/A3B related mutations maybe associated with poor prognosis and therapeutic resistance in cancers. We are developing ssDNA based A3A/A3B inhibitors as ssDNA is the natural substrate for their catalytic reaction. A transition state analogue, 2'-deoxy-zebularine, was incorporated into the target ssDNA sequence of A3A/A3B, and yielded promising affinity and inhibitory effect. We are currently testing modification of nucleotides in order to resist degradation in cells.

APOBEC3G（A3G）和APOBEC3H（A3H）是单链DNA胞嘧啶脱氨酶，可以通过突变HIV-1基因组来限制HIV-1感染。 HIV-1开发了一种反式机制，通过该机制，病毒体感染因子（VIF）通过泛素 - 蛋白酶体途径导致A3G/A3H的降解。我们的最终目标是生成抑制A3G/A3H降解的小化合物。以前，我们已经确定了A3蛋白域的NMR和晶体结构，包括VIF结合域和催化域。此外，我们确定了A3催化结构域-SSDNA复合物的共结晶结构。在2021 - 2022年期间，我们开发了一种技术，可以选择性地与A3G复合物，并分配了VIF的NMR信号，以鉴定具有A3G的接口中的氨基酸残基（JMR，JMR，20222年）。我们通过使用Cryoem来确定A3H-VIF E3泛素连接酶复合物的结构取得了进展，这将提供由抑制复合物形成的小化合物靶向的表位。 APOBEC3A（A3A）和APOBEC3B（A3B）蛋白的失调贡献了大约75％的癌症类型的DNA突变的主要内源性来源，所有分析的癌症的50％。 A3A/A3B蛋白可以单独引起DNA突变，也可以作为对癌症疗法的反应，而癌症疗法可以驱动癌症的进化，而A3A/A3B相关的突变可能与癌症的预后和预后不良和治疗性有关。我们正在开发基于ssDNA的A3A/A3B抑制剂，因为ssDNA是其催化反应的天然底物。将过渡态类似物，2'-脱氧 - Zebularine纳入了A3A/A3B的靶标序列中，并产生了有希望的亲和力和抑制作用。我们目前正在测试核苷酸的修饰，以抵抗细胞中的降解。