MOLECULAR ANALYSIS OF FANCONI'S ANEMIA C PROTEIN

这个项目的长期目标是阐明 “逻辑”与参与造血细胞基因组稳定性的分子 通过对Fanconi贫血(FA)的研究。现在已经克隆了三个FA基因， 调查员准备询问有关该组织的准确问题 和他们的蛋白质产品的功能。虽然FA的细胞表型 在药物敏感通路中牵涉到这些蛋白质作为 基因组的稳定性，它们的分子功能仍然不完全清楚。 FANCC通过其与细胞的相互作用而在细胞解毒中发挥作用 NADPH细胞色素P-450还原酶(RED)和DNA损伤前的调节步骤。 FANCA是过氧化物酶的同源物，与FANCG相互作用，并在 原子核。利用细胞培养酵母和小鼠模型，研究人员建议 检验细胞质FANCC-RED和核FANCA-FANCG的假设 复合体在其各自的细胞内执行解毒功能 车厢。因此，调查者将(I)表征该表达式 小鼠胚胎发育过程中FA基因产物的模式 通过原位和生化策略进行造血细胞和生殖细胞发育； (Ii)确定FA蛋白的寡聚体结构和调控；(Iii) 使用遗传策略定位FA蛋白的功能以预防或 DNA损伤后步骤，并在此背景下测试FANCA的功能 过氧化物酶结构域；以及(Iv)分离调节FANCC-red途径的基因， 并描述了这一途径与由其调控的途径的关系 FANCA-FANCG。我们结合遗传、细胞和生物化学的方法应该 结果对FA基因的调控和功能有了全面的认识 产品。除了提高我们对基本机制的理解之外 与造血有关的细胞解毒和染色体稳定性， 操纵由FA基因控制的药物敏感通路将提供 白血病或固体化疗增敏的新翻译机会 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.