FANCM in repair of stalled replication forks

FANCM 修复停滞的复制叉

• 批准号: 9363243
• 负责人: Ralph Scully
• 金额: $ 39.57万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-28 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9363243
• 关键词: Acute Myelocytic Leukemia; Affect; Alpha Cell; Anemia; BARD1 gene; BRCA1 gene; BRCA2 gene; Cells; Childhood; Clinical; Collaborations; Complement; Complex; Congenital Abnormality; DNA; DNA Damage; DNA Repair; DNA Repair Pathway; DNA Replication Damage; DNA Structure; DNA replication fork; DNA-Directed DNA Polymerase; Development; Disease; Enzymes; Escherichia coli; Fanconi Anemia pathway; Fanconi' s Anemia; Gene Conversion; Generic Drugs; Genes; Genomic Instability; Goals; Health; Hereditary Breast Carcinoma; Hereditary Breast and Ovarian Cancer Syndrome; Human; Incidence; Interruption; Link; Malignant Neoplasms; Mammalian Cell; Massachusetts; Mediating; Metabolism; Modeling; Molecular; Motor Activity; Mus; Mutate; Mutation; Nature; Organ; Outcome; Pancytopenia; Pathway interactions; Play; Predisposition; Price; Process; Proteins; Recruitment Activity; Reporter; Role; Site; Solid Neoplasm; Stress; Switching Complex; Time; United States National Institutes of Health; Ursidae Family; Work; Yeasts; cancer cell; cancer genome; cancer risk; cancer therapy; embryonic stem cell; endodeoxyribonuclease SceI; genetic analysis; helicase; homologous recombination; human disease; insight; malignant breast neoplasm; medical schools; mutant; new therapeutic target; novel; nuclease; prevent; repaired; response; success; targeted cancer therapy; tool

Replication fork stalling at sites of abnormal DNA structure is a recognized cause of genomic instability. Increased replication fork stalling (“replication stress”) is a common feature of cancer cells, suggesting that defective processing of the stalled fork is a common mechanism of genomic instability in cancer. The Fanconi Anemia (FA) pathway has a major role in the metabolism and repair of stalled replication forks. FA is a rare, autosomal recessive (or X-linked) disease caused by inactivation of any one of several FA genes. The clinical manifestations of FA include childhood anemia and progressive bone marrow failure, together with short stature and congenital defects affecting a wide variety of organs. The risk of cancer, including solid tumors, is elevated, with particularly high incidence of acute myelogenous leukemia. The gene encoding an early responder of FA pathway, FANCM, is found mutated in some breast cancers. The FA pathway overlaps functionally with the BRCA pathway of hereditary breast/ovarian cancer predisposition—a critical regulator of homologous recombination. The FA pathway is also activated by replication stress, indicating a general role for the FA genes in human cancer and in many other diseases. Thus, deciphering the mechanisms of action of the FA pathway has broad significance for human health. We recently adapted the Escherichia coli Tus/Ter replication fork arrest complex for use in mammalian cells and have used it to quantify both error-free and error-prone homologous recombination induced by a mammalian chromosomal replication fork block. More recently, we identified a novel aberrant repair product of replication fork arrest in mammalian cells, in which small (<10 kb) microhomology-mediated tandem duplications form at the site of replication arrest. FANCM plays a crucial role in suppressing these aberrant repair products at stalled forks. In work proposed here, we will use novel tools, recently developed by the Scully lab, to analyze how FANCM regulates homologous recombination at stalled replication forks. We will identify the mechanisms by which FANCM suppresses tandem duplication at stalled forks. Success in this work will lead to the identification of new targets for therapy in cancer and other human diseases.

复制叉在 DNA 结构异常位点停滞是基因组不稳定的公认原因。 复制叉停滞（“复制应激”）增加是癌细胞的一个共同特征，这表明 停滞叉的加工缺陷是癌症基因组不稳定的常见机制。范科尼 贫血（FA）途径在停滞复制叉的代谢和修复中发挥着重要作用。 FA是一种罕见的、 由几种 FA 基因中的任何一种失活引起的常染色体隐性遗传（或 X 连锁）疾病。临床上 FA 的表现包括儿童期贫血和进行性骨髓衰竭，以及短暂的 影响多种器官的身材和先天缺陷。患癌症（包括实体瘤）的风险是 升高，其中急性粒细胞白血病的发病率特别高。编码早期基因 FA 途径的反应者 FANCM 在某些乳腺癌中被发现发生突变。 FA 途径重叠 与遗传性乳腺癌/卵巢癌易感性的 BRCA 通路有功能，是乳腺癌/卵巢癌的关键调节因子 同源重组。 FA 通路也会被复制应激激活，表明 FA 通路的一般作用 人类癌症和许多其他疾病中的 FA 基因。因此，破译其作用机制 FA途径对人类健康具有广泛的意义。我们最近改编了大肠杆菌 Tus/Ter 复制叉阻滞复合物用于哺乳动物细胞，并用它来定量无误和 由哺乳动物染色体复制叉块诱导的易错同源重组。更多的 最近，我们在哺乳动物细胞中发现了一种新的复制叉停滞异常修复产物，其中 小的（<10 kb）微同源介导的串联重复在复制停滞位点形成。 FANCM 在抑制停顿叉处的这些异常修复产物方面发挥着至关重要的作用。在这里提出的工作中，我们 将使用 Scully 实验室最近开发的新颖工具来分析 FANCM 如何调节同源 在停滞的复制叉处重组。我们将确定 FANCM 抑制的机制 停滞叉处的串联复制。这项工作的成功将有助于确定新的治疗靶点 在癌症和其他人类疾病中。