MOLECULAR ANALYSIS OF FANCONI'S ANEMIA C PROTEIN

FANCONI 贫血 C 蛋白的分子分析

• 批准号: 6128945
• 负责人: HAGOP YOUSSOUFIAN
• 金额: $ 29.9万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-07-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6128945
• 关键词: DNA damage; biological signal transduction; chimeric proteins; congenital aplastic anemia; cytochrome P450; detoxification; embryogenesis; gel electrophoresis; gene expression; gene targeting; hematopoiesis; laboratory mouse; molecular pathology; protein protein interaction; protein structure function; southern blotting; tissue /cell culture

The long term goals of this project are to elucidate both the 'logic' and molecules involved in the genomic stability of hematopoietic cells through studies of Fanconi anemia (FA). Three FA genes have now been cloned, and the investigator is poised to ask precise questions about the organization and function of their protein products. Although the cellular phenotype of FA implicates these proteins in drug-sensitive pathways as "gatekeepers" of genomic stability, their molecular functions remain incompletely understood. FANCC has a role in cellular detoxification by virtue of its interaction with NADPH cytochrome P-450 reductase (RED) and regulation of a pre-DNA damage step. FANCA is homologous to peroxidases, interacts with FANCG, and functions in the nucleus. Using cell culture yeast and mouse models, the investigator proposes to test the hypothesis that cytoplasmic FANCC-RED and nuclear FANCA-FANCG complexes perform detoxification functions in their respective cellular compartments. Thus, the investigator will (I) characterize the expression patterns of FA gene products during mouse embryogenesis, including hematopoietic and germ cell development, by in situ and biochemical strategies; (II) determine the oligomeric structure and regulation of FA proteins; (III) use genetic strategies to IocaIize the function of FA proteins to pre- or post-DNA damage steps, and, in this context, test the function of the FANCA peroxidase domain; and (IV) isolate genes that regulate the FANCC-RED pathway, and characterize the relationship of this pathway to that regulated by FANCA-FANCG. Our combined genetic, cellular and biochemical approaches should result in a comprehensive view of the regulation and function of FA gene products. Aside from improving our understanding of fundamental mechanisms of cellular detoxification and chromosomal stability relevant to hematopoiesis, the manipulation of drug-sensitive pathways controlled by FA genes will provide novel translational opportunities for chemosensitization of leukemias or solid tumors to bifunctional cross-linkers.

该项目的长期目标是阐明 参与造血细胞基因组稳定性的“逻辑”和分子 通过Fanconi贫血（FA）的研究。现在已经克隆了三个FA基因， 调查员准备提出有关组织的精确问题 和蛋白质产品的功能。虽然FA的细胞表型 将这些蛋白在药物敏感途径中含义为 基因组稳定性，它们的分子功能仍然不完全理解。 FANCC通过与 NADPH细胞色素P-450还原酶（红色）和DNA损伤步骤的调节。 Fanca与过氧化物酶是同源的，与Fancg相互作用，并在 核。使用细胞培养酵母和小鼠模型，研究者提出了 为了检验细胞质fancc-red和核芬卡 - 芬奇的假设 复合物在其各自的细胞中执行排毒功能 车厢。因此，研究者将（i）表征表达式 小鼠胚胎发生过程中FA基因产物的模式，包括 造血和生殖细胞发育，通​​过原位和生化策略； （ii）确定FA蛋白的寡聚结构和调节； （iii） 利用遗传策略来使FA蛋白的功能在前或 DNA损伤步骤，在这种情况下，测试FANCA的功能 过氧化物酶结构域； （iv）调节FANCC-RED途径的分离基因， 并表征该途径与所规定的途径的关系 Fanca-Fancg。我们的遗传，细胞和生化方法应该 导致对FA基因的调节和功能的全面视图 产品。除了提高我们对基本机制的理解 与造血相关的细胞排毒和染色体稳定性， FA基因控制的药物敏感途径的操纵将提供 白血病或固体化学敏化的新型翻译机会 肿瘤到双功能交联。