MOLECULAR ANALYSIS OF FANCONI'S ANEMIA C PROTEIN

本项目的长期目标是阐明 造血细胞基因组稳定性中的“逻辑”和分子 范可尼贫血（FA）目前已经克隆了三个FA基因， 调查员准备问一些关于组织的问题 及其蛋白质产物的功能。虽然FA的细胞表型 暗示这些蛋白质在药物敏感途径中作为药物敏感的“看门人”， 基因组的稳定性，其分子功能仍然不完全了解。 FANCC通过其与以下物质的相互作用而在细胞解毒中起作用： NADPH细胞色素P-450还原酶（RED）和前DNA损伤步骤的调节。 FANCA与过氧化物酶同源，与FANCG相互作用，并在细胞内起作用。 原子核使用细胞培养酵母和小鼠模型，研究者提出， 为了验证细胞质FANCC-RED和细胞核FANCA-FANCG 复合物在它们各自的细胞中执行解毒功能， 隔间因此，研究人员将（I）表征表达 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蛋白的寡聚体结构和调节;（III） 用遗传学的方法来定位FA蛋白的功能， DNA损伤后的步骤，并在这种情况下，测试FANCA的功能， 过氧化物酶结构域;和（IV）分离调节FANCC-RED途径的基因， 并描述该途径与受 FANCA-FANCG。我们结合遗传、细胞和生物化学方法， 从而对FA基因的调控和功能有了一个全面的认识 产品.除了提高我们对基本机制的理解， 与造血相关的细胞解毒和染色体稳定性， 对由FA基因控制的药物敏感途径的操纵将提供 白血病或实体瘤化疗增敏的新的转化机会 肿瘤到双功能交联剂。

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.