Structural Biology of XPB and XPD Helicases

XPB 和 XPD 解旋酶的结构生物学

• 批准号: 8212285
• 负责人: John A. Tainer
• 金额: $ 32.82万
• 依托单位: UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8212285
• 关键词: Address; Aging; Amino Acids; Architecture; Back; Binding; Biochemical; Biological; Biology; Blinded; Cell Death; Chemistry; Child; Clinic; Cockayne Syndrome; Code; Complex; DNA; DNA Damage; DNA Repair; DNA Structure; Defect; Disease; ERCC3 gene; Employee Strikes; Event; Future; Gene Mutation; Genes; Genetic Code; Genetic Transcription; Germ-Line Mutation; Goals; Grant; Health; Homologous Gene; Human; Inherited; Iron; Lead; Letters; Malignant Neoplasms; Mediating; Modeling; Molecular; Molecular Conformation; Moon; Mutation; Names; Nature; Neurologic; Nucleotide Excision Repair; Nucleotides; Outcome; Oxidation-Reduction; Pathway interactions; Patients; Phenotype; Play; Predisposition; Premature aging syndrome; Process; Proteins; RNA Polymerase II; Repair Complex; Research; Resolution; Role; Skin Cancer; Solutions; Specificity; Structural Biochemistry; Structure; Sulfur; Testing; The Sun; Therapeutic; Transcription-Coupled Repair; Trichothiodystrophy; Xeroderma Pigmentosum; base; cancer cell; cancer risk; clinical phenotype; cofactor; disease phenotype; disease-causing mutation; flexibility; helicase; human disease; insight; member; mutant; protein protein interaction; repaired; research study; stem; structural biology; transcription factor TFIIH; Address; Aging; Amino Acids; Architecture; Back; Binding; Biochemical; Biological; Biology; Blinded; Cell Death; Chemistry; Child; Clinic; Cockayne Syndrome; Code; Complex; DNA; DNA Damage; DNA Repair; DNA Structure; Defect; Disease; ERCC3 gene; Employee Strikes; Event; Future; Gene Mutation; Genes; Genetic Code; Genetic Transcription; Germ-Line Mutation; Goals; Grant; Health; Homologous Gene; Human; Inherited; Iron; Lead; Letters; Malignant Neoplasms; Mediating; Modeling; Molecular; Molecular Conformation; Moon; Mutation; Names; Nature; Neurologic; Nucleotide Excision Repair; Nucleotides; Outcome; Oxidation-Reduction; Pathway interactions; Patients; Phenotype; Play; Predisposition; Premature aging syndrome; Process; Proteins; RNA Polymerase II; Repair Complex; Research; Resolution; Role; Skin Cancer; Solutions; Specificity; Structural Biochemistry; Structure; Sulfur; Testing; The Sun; Therapeutic; Transcription-Coupled Repair; Trichothiodystrophy; Xeroderma Pigmentosum; base; cancer cell; cancer risk; clinical phenotype; cofactor; disease phenotype; disease-causing mutation; flexibility; helicase; human disease; insight; member; mutant; protein protein interaction; repaired; research study; stem; structural biology; transcription factor TFIIH

DESCRIPTION (provided by applicant): Hereditary mutations in the DNA helicases XPB and XPD lead to human diseases with different phenotypes reflecting increased cancers or aging: xeroderma pigmentosum (XP), XP combined with Cockayne syndrome (CS), and trichothiodystrophy (TTD). These diseases reflect the disruption of different cellular pathways: nucleotide-excision repair (NER), transcription-coupled repair (TCR), or transcription. In humans, XPB and XPD helicases are part of the ten subunit TFIIH transcription/repair complex, but disease-causing mutations cluster in XPB and particularly XPD rather than in the other TFIIH proteins, excepting TFB5, so these XP helicases appear key to controlling coordination of transcription and repair. We aim to understand the molecular features underlying the specificity, activity, conformational controls and pathway coordination by the XPB and XPD helicases. Our hypothesis is that well-defined architectures, conformational states, and molecular interfaces of XPB and XPD helicases provide critical controls for transcription, NER, and TCR. We have shown that characterizations of these features and their disruption by disease-causing mutations provide a molecular basis to directly connect the inherited gene mutations to disease phenotypes. Building on our crystal structures of XPB and XPD, we propose to integrate structural and biophysical experiments including small angle x-ray scattering to define conformations and complexes in solution with biochemical and biological experiments to determine structures of disease-relevant mutants, protein-DNA complexes, and define key interactions for their activities. The anticipated outcome of the proposed cross-disciplinary experiments is a molecular picture of the protein-DNA complexes, protein-protein interactions and functional states that orchestrate transcription and repair events mediated by XPB and XPD as components of TFIIH. These results will help provide a detailed molecular understanding of the processes that underlie the cancer and cell death disease phenotypes associated with XP, XP/CS, and TTD patient mutations.

描述（由申请人提供）：DNA解旋酶XPB和XPD中的遗传突变导致人类疾病，反映出癌症或衰老的表型不同：Xeroderma cipmentosum（XP），XP与Cockayne综合征（CS）和Trichothiothiodododystrophy（TTD）结合在一起。这些疾病反映了不同细胞途径的破坏：核苷酸 - 脱孔修复（NER），转录耦合修复（TCR）或转录。在人类中，XPB和XPD解放酶是十个亚基TFIIH转录/修复复合物的一部分，但是引起疾病的突变集群在XPB，尤其是XPD中，而不是在其他TFB5蛋白中，除了TFB5以外的其他TFIIH蛋白质，因此这些XP Helicass似乎是控制转录和维修均衡的关键。我们旨在了解XPB和XPD解旋酶的特异性，活性，构象控制和途径协调的基础分子特征。我们的假设是，XPB和XPD解酶的定义明确的结构，构象状态和分子界面为转录，NER和TCR提供了关键的控制。我们已经表明，这些特征的特征及其因疾病突变而被破坏提供了分子基础，可以将遗传基因突变与疾病表型直接连接起来。在我们的XPB和XPD的晶体结构的基础上，我们建议整合结构和生物物理实验，包括小角度X射线散射，以定义与生物化学和生物学实验溶液中的构象和复合物，以确定疾病相关的突变体的结构，蛋白质-DNA复合物，并为其活性定义键盘相互作用。拟议的跨学科实验的预期结果是蛋白质-DNA复合物，蛋白质 - 蛋白质相互作用和功能状态的分子图，这些状态策划了由XPB和XPD作为TFIIH组成部分的转录和修复事件。这些结果将有助于提供对与XP，XP/CS和TTD患者突变相关的癌症和细胞死亡疾病表型基础的过程的详细分子理解。