Model Genetic Systems to Study DNA Repair

建立遗传系统模型来研究 DNA 修复

• 批准号: 7732313
• 负责人: Robert Brosh
• 金额: $ 18.23万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7732313
• 关键词: Alkylating Agents; Biological; Biological Models; Cells; Chromosomal Stability; Critical Pathways; DNA; DNA Repair; DNA Repair Pathway; DNA Structure; DNA biosynthesis; Diagnosis; Disease; Embryo; Evolution; Fanconi' s Anemia; Fibroblasts; G-Quartets; Genetic; Genetic Models; Genome; Genome Stability; Genomic Instability; Helicase Gene; Hereditary Disease; Homeostasis; Human; Human Cell Line; Individual; Knockout Mice; Link; Maintenance; Malignant Neoplasms; Molecular; Mus; Mutation; Orthologous Gene; Pathway interactions; Premature aging syndrome; RECQL gene; RNA Interference; Resistance; Role; Stress; System; WRN gene; Werner Syndrome; Work; Yeasts; base; helicase; in vivo; insight; novel; repaired; small molecule

Using an approach known as RNA interference, we have specifically depleted human cells of the FANCJ DNA helicase and characterized the sensitivity of the cells to a small molecule compound that stabilizes G-quadruplex DNA structures. This work enabled us to elucidate a novel functionof the FANCJ helicase in the manintenace of chromosomal stability. Since individuals carrying homozygous mutations in the FANCJ helicase gene have a genetic disorder known as Fanconi Anemia characterized by genomic instability and cancer, we believe our results shed new insights to the cellular pathways of FANCJ that serve to counter replciational stress due to alterante DNA structures such as G-quadruplexes that arise in vivo. We have also employed yeast as a model genetic system to study the role of the Werner syndrome helicase in DNA replciation and repair. This work has enabled us to characterize the catalytic requirements of WRN in a defined genetic DNA repair pathway that operates to provide cellular resistance to alkylating agents which impose replicational stress. Lastly, we have utilized mouse as a model genetic system to characterize the role of a helicase ortholog (RECQ1) found in humans whose biological significance is not well understood. Characterization of th eprimary mouse embryonic fibroblasts from RECQ1 knockout mice has revealed that the RECQ1 helicase has unique and important roles in genomic stability maintenance.

使用一种称为RNA干扰的方法，我们特别耗尽了FANCJ DNA解旋酶的人类细胞，并表征了细胞对稳定G- Qu-Quadruplex DNA结构的小分子化合物的敏感性。 这项工作使我们能够在染色体稳定性的Manintenace中阐明FANCJ解旋酶的新功能。 由于在FANCJ解放酶基因中携带纯合突变的个体具有一种以基因组不稳定性和癌症为特征的遗传疾病，因此我们相信我们的结果为FANCH的细胞途径提供了新的见解，该途径是由诸如g-Quarplexees诸如Arise arise vivo的anterante dna结构所引起的替代DNA结构所引起的。 我们还利用酵母作为模型遗传系统来研究Werner综合征解旋酶在DNA反式围和修复中的作用。 这项工作使我们能够表征WRN在定义的遗传DNA修复途径中的催化需求，该途径可为烷基化剂提供施加复制应激的烷基化剂的细胞耐药性。 最后，我们利用小鼠作为模型遗传系统来表征在人类中发现的解旋酶直系同源物（RECQ1）的作用，该人的生物学意义尚不清楚。 RECQ1基因敲除小鼠的电子胚胎胚胎成纤维细胞的表征表明，RECQ1解旋酶在基因组稳定性维持中具有独特而重要的作用。