Molecular Pathogenesis of Fanconi Anemia

范可尼贫血的分子发病机制

基本信息

批准号：
10083752
负责人：
ALAN D. D'ANDREA
金额：
$ 43.14万
依托单位：
DANA-FARBER CANCER INST
依托单位国家：
美国
项目类别：
财政年份：
1994
资助国家：
美国
起止时间：
1994-08-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10083752
关键词：
ATP phosphohydrolase Acute Myelocytic Leukemia Affect Anemia BRCA2 gene Binding Binding Proteins Bypass C-terminal Characteristics Chromosomal Instability Collaborations Complex Congenital Abnormality DNA DNA Crosslinking Agent DNA Damage DNA Interstrand Crosslinking DNA Repair DNA Repair Pathway DNA crosslink Data Development Disease Dysmyelopoietic Syndromes Elements Event Excision FANCD2 protein Fanconi Anemia pathway Fanconi Anemia-BRCA Pathway Fanconi anemia protein Fanconi&apos s Anemia Genes Genetic Genetic Diseases Genomic Instability Grant Hypersensitivity Inherited Knock-out Knockout Mice Laboratories Malignant Neoplasms Mass Spectrum Analysis Mediating Mitomycin C Molecular Molecular Conformation Multiprotein Complexes Mutation Nonhomologous DNA End Joining Pancytopenia Pathogenesis Pathway interactions Patients Phenotype Play Polymerase Population Predisposition Process Proteins Rare Diseases Regulation Resistance Role Site cancer predisposition crosslink cytotoxic endonuclease inhibitor/antagonist knock-down malignant breast neoplasm mouse model neoplastic cell novel nuclease overexpression p53-binding protein 1 recessive genetic trait recruit repair function repaired replication stress scaffold

项目摘要

PROJECT SUMMARY Fanconi Anemia (FA) is a rare autosomal recessive genetic disease characterized by congenital defects, bone marrow failure, cancer predisposition, and cellular hypersensitivity to DNA crosslinking agents (ICLs). FA is caused by mutations in one of 23 genes whose protein products collaborate in a pathway required for removing cytotoxic interstrand crosslinks (ICLs) from DNA. The D'Andrea laboratory has identified many of the molecular players and processes in the FA/BRCA pathway. In collaboration with the Soulier laboratory, we recently identified REV7 as the FA gene, FANCV. This was a key finding, since REV7/FANCV is emerging as a critical protein affecting several different DNA repair processes, by engaging with different binding partners through its C-terminal “seatbelt” domain. When the seatbelt of REV7 is bound to SHLD3, it functions upstream in NHEJ. When the seatbelt of REV7 is bound to REV3, it functions as a translesion (TLS) polymerase, Polζ, required for bypassing an unhooked DNA crosslink, generated by the upstream FA proteins. Thus, REV7 deficiency, like the deficiency of other FA proteins, causes sensitivity to Mitomycin C (MMC), an ICL-inducing agent. Through preliminary studies for this grant, we identified novel REV7 interactors by IP-MS. Interestingly, we identified the binding partner, TRIP13, a novel ATPase which flips REV7 into an open conformation and releases REV3 and inactivates the FA/BRCA pathway. We also identified the REV7-binding protein SHLD2/FAM35A. Knockdown of this protein, like REV7 knockdown, disrupts the FA/BRCA pathway. The Specific Aims for the next five years of this grant are 1) to determine the role of the ATPase TRIP13 in the suppression of the FA/BRCA Pathway 2) to determine the role of the REV7/FANCV protein and its binding partner SHLD2/FAM35A in the activation of the FA/BRCA pathway 3) to employ knockout mouse models for REV7/FANCV and TRIP13 to evaluate the regulation of the FA/BRCA pathway.

项目概要范可尼贫血（FA）是一种罕见的常染色体隐性遗传病，其特征是先天性缺陷、骨骨髓衰竭、癌症易感性和细胞对 DNA 交联剂 (ICL) 过敏。由 23 个基因之一的突变引起，这些基因的蛋白质产物在一条所需的途径中协同作用 D'Andrea 实验室已鉴定出许多 DNA 中的细胞毒性链间交联 (ICL)。 FA/BRCA 途径中的分子参与者和过程我们与 Soulier 实验室合作。最近将 REV7 确定为 FA 基因 FANCV，这是一项重要发现，因为 REV7/FANCV 正在作为 FA 基因出现。一种关键蛋白质，通过与不同的结合伙伴结合，影响几种不同的 DNA 修复过程通过其C端“安全带”结构域当REV7的安全带与SHLD3结合时，它发挥上游作用。在 NHEJ 中，当 REV7 的安全带与 REV3 结合时，它充当跨损伤 (TLS) 聚合酶 Pol z，绕过由上游 FA 蛋白产生的脱钩 DNA 交联所需的。与其他 FA 蛋白的缺乏一样，缺乏会导致对丝裂霉素 C (MMC) 的敏感性，丝裂霉素 C 是一种 ICL 诱导剂。通过本次资助的初步研究，我们通过 IP-MS 鉴定了新型 REV7 相互作用分子。我们确定了结合伴侣 TRIP13，这是一种新型 ATP 酶，可将 REV7 翻转为开放构象，释放 REV3 并使 FA/BRCA 途径失活我们还鉴定了 REV7 结合蛋白。 SHLD2/FAM35A 的敲除（如 REV7 敲除）会破坏 FA/BRCA 通路。这笔赠款未来五年的具体目标是 1) 确定 ATPase TRIP13 在抑制 FA/BRCA 途径 2) 以确定 REV7/FANCV 蛋白的作用及其结合合作伙伴 SHLD2/FAM35A 激活 FA/BRCA 途径 3) 采用敲除小鼠模型 REV7/FANCV 和 TRIP13 用于评估 FA/BRCA 通路的调节。