Functional Genomics

功能基因组学

• 批准号: 8494583
• 负责人: RAYMOND J MONNAT
• 金额: $ 25.39万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8494583
• 关键词: Adult; Affinity Chromatography; Alleles; Biological Assay; Biology; Bloom Syndrome; Cell Proliferation; Cells; Chemicals; Chemotherapy-Oncologic Procedure; DNA; DNA Damage; DNA biosynthesis; Disease; Epithelial; Gene Proteins; Genetic; Genetic Recombination; Genome Stability; Goals; Human; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Mediating; Metabolic; Metabolic Pathway; Metabolism; Microfluidics; Mutation; Pathway interactions; Pharmaceutical Preparations; Post-Translational Modification Site; Post-Translational Protein Processing; Predisposition; Proteins; Proteomics; RECQL4 gene; RecQ protein; Research; Research Personnel; Resolution; Role; Rothmund-Thomson syndrome; Site; Syndrome; Werner Syndrome; Work; cancer risk; chemotherapeutic agent; chemotherapy; clinically relevant; functional genomics; helicase; human WRN protein; human disease; improved; insight; killings; mutant; neoplastic cell; oncology; programs; protein function; repaired; response; single molecule; small hairpin RNA; small molecule; tumor

The goals of Project 2 are to develop a detailed picture of human disease-associated RecQ helicase protein function, and to use this information to delineate the role of these RecQ helicases in human tumor cell proliferation, survival and the response to chemotherapy. In research to date we have focused on functional analyses of the human Werner syndrome protein WRN. In the proposed research we extend these analyses to encompass all three disease-associated human RecQ helicases, the WRN, BLM and RECQL4 proteins associated with the heritable deficiency/genetic instability/cancer predisposition syndromes Werner syndrome (WRN), Bloom syndrome (BLM) and Rothmund-Thomson (RTS) syndrome, respectively. Aim 1. Develop a comprehensive proteomic description of the human disease-associated WRN, BlM and RECQl4 human RecQ helicases. We will identify functionally important RecQ helicase protein associations and post-translational modifications (PTM's) by using quantitative affinity purification mass spectrometry, shRNA-mediated epistatic interaction mapping and functional analyses of post-translational modification (PTM) site mutants. Aim 2. Determine functional requirement for WRN, BlM and RECQL4 in DNA metabolism. We will determine the role of each of these RecQ helicases in DNA replication, translesion DNA synthesis (TLS), homology-dependent recombination (HDR) and DNA break repair using high resolution assays. Aim 3. Determine functional redundancy among WRN, BlM and RECQL4 in DNA metabolism, and clinically relevant synthetic interactions with additional genes/proteins, drugs or small molecules. We will identify unique and redundant functions of the 'disease-associated human RecQ helicases, and synthetic interactions between disease-associated human RecQ helicases, their interacting proteins, and chemotherapeutic agents, drugs or small molecules that can beused to selectively kill RecQ-deficient human tumors.

项目2的目标是开发与人类疾病相关的RECQ解旋酶蛋白功能的详细图片，并使用此信息来描述这些RECQ解旋酶在人肿瘤细胞增殖，生存和对化疗的反应中的作用。在迄今为止的研究中，我们专注于人类Werner综合征蛋白WRN的功能分析。在拟议的研究中，我们扩展了这些分析，以涵盖所有与疾病相关的人类RECQ解旋酶，即WRN，BLM和RECQL4蛋白与可遗传的缺乏/遗传不稳定性/癌症倾向综合征有关 综合征（WRN），Bloom综合征（BLM）和Rothmund-Thomson（RTS）综合征。 目标1。开发与人类疾病相关的WRN，BLM和RECQL4人RECQ解旋酶的全面蛋白质组学描述。我们将通过使用定量亲和力纯化质谱法，SHRNA介导的上皮相互作用映射和翻译后变性后修饰（PTM）位点突变体的功能分析来鉴定具有功能上重要的RECQ解旋蛋白蛋白关联和翻译后修饰（PTM）。 AIM 2。确定DNA代谢中WRN，BLM和RECQL4的功能需求。我们将使用高分辨率测定法确定每个RECQ解旋酶在DNA复制，Translesion DNA合成（TLS），同源依赖性重组（HDR）和DNA断裂修复中的作用。 AIM 3。确定DNA代谢中WRN，BLM和RECQL4之间的功能冗余，以及与其他基因/蛋白质，药物或小分子的临床相关合成相互作用。我们将确定与疾病相关的人RECQ解旋酶的独特和冗余功能，以及与疾病相关的人类RECQ解旋酶，它们相互作用的蛋白质以及化学治疗剂，药物或小分子之间的合成相互作用 人类肿瘤。