An organismal approach to dissecting the mechanism of DSB repair pathway choice

剖析 DSB 修复途径选择机制的有机方法

• 批准号: 9021045
• 负责人: Sarit Smolikove
• 金额: $ 8.66万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-15 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9021045
• 关键词: Accounting; Address; Affect; Alleles; Animal Model; Biochemical; Biological Assay; Biological Models; Caenorhabditis elegans; Cancer Patient; Candidate Disease Gene; Catalytic Domain; Cell Cycle Proteins; Cell Cycle Regulation; Cell Death; Cell Nucleus; Cell physiology; Cells; Cellular biology; Characteristics; Chromatin; Chromosomal translocation; Chromosome Structures; Chromosome abnormality; Complement; Complex; DNA; DNA Damage; DNA Double Strand Break; DNA Repair; DNA Sequence Alteration; Data; Development; Diagnosis; Double Strand Break Repair; Excision; Future; Gene Expression Regulation; Gene Fusion; Gene Structure; Genes; Genetic; Genetic Epistasis; Genetic Screening; Genome; Genomic Instability; Goals; Human; Lead; Life; Malignant Neoplasms; Mammals; Measures; Meiosis; Methods; Modeling; Molecular; Molecular Genetics; Molecular Mechanisms of Action; Morbidity - disease rate; Nonhomologous DNA End Joining; Organism; Outcome; Participant; Pathway interactions; Pharmaceutical Preparations; Play; Prevention; Process; Property; Proteins; Public Health; RNA Interference; Radiation therapy; Reagent; Regulation; Replication Error; Research; Role; Single-Stranded DNA; System; Testing; Time; Translational Research; V(D)J Recombination; Whole Organism; Yeasts; base; cancer cell; cancer therapy; chromatin modification; cost; design; ds-DNA; gene conservation; gene discovery; gene repair; genome integrity; homologous recombination; improved; insertion/deletion mutation; insight; mutant; novel; prevent; programs; protein complex; public health relevance; repaired; therapy design; tumor; tumor progression; yeast two hybrid system

DESCRIPTION (provided by applicant): Project Summary A double stand break is a type of DNA damage which involves the severing of one DNA molecule into two parts. Unrepaired breaks are harmful to living organisms since they lead to cell death. However, cells that do repair their breaks can also suffer detrimental outcomes if the repair is associated with DNA alteration. Repair pathways can be error free, restoring the DNA to its pristine condition, or error-prone, leading to mistakes in DNA repair. The latter may lead to chromosomal abnormalities frequently associated with changes in gene structure and regulation. The effects of these errors on chromosome structure and gene regulation often promote tumors and are frequently found in cells of cancer patient. A low level of DNA breaks occur due to normal cellular process and some are environmentally induced (for example, radiation therapy and/or chemotherapeutic drugs). If this damage is not properly repaired, it can contribute to cancer reoccurrence. Understanding processes involved in how a particular DNA repair mechanism is selected will be fundamental for determining how cancer progresses as well as for cancer treatment. The long-term goal of this research is to identify mechanisms involved in selecting an error-free repair vs. an error-prone repair pathway. This proposal focuses on identifying proteins and mechanisms involved in this choice. Studies are proposed to understand how processing of double stand break affects such a decision. We will use C. elegans, a multi- cellular animal model system which allows us to preform genetic assays for DNA repair in whole organism, at a low cost and with efficient time scale. Using a newly identified mutant in a key repair gene, we will examine DNA processing independently of DNA damage in the C. elegans germline. Furthermore, we will identify new mechanisms directly regulating a key protein involved in this processing. To complement this analysis, we will study the germline as a model for the cell-cycle control of DNA processing by designing a system for targeting DNA damage to particular regions of the germline. The conservation of genes and pathways between this animal model and humans will serve as efficient approach for discovery of genes in humans affecting the formation and progression of cancer. Understanding how accurate versus error-prone pathways are selected will be essential for developing anti-cancer therapies utilizing DNA damage to destroy cancer cells without harming healthy non-cancerous cells.

描述（由申请人提供）： 双链断裂是一种DNA损伤，涉及一个DNA分子被切断成两部分。未修复的断裂对生物体有害，因为它们会导致细胞死亡。然而，如果修复与DNA改变有关，修复断裂的细胞也会遭受有害的结果。修复途径可以是无错误的，将DNA恢复到其原始状态，或容易出错，导致DNA修复错误。的 后者可能导致染色体异常，通常与基因结构和调节的变化有关。这些错误对染色体结构和基因调控的影响通常会促进肿瘤的发生，并且经常在癌症患者的细胞中发现。低水平的DNA断裂是由于正常的细胞过程而发生的，有些是环境诱导的（例如，放射治疗和/或化疗药物）。如果这种损伤没有得到适当的修复，它可能会导致癌症复发。了解如何选择特定DNA修复机制所涉及的过程将是确定癌症进展以及癌症治疗的基础。这项研究的长期目标是确定参与选择无错误修复与易错修复途径的机制。这项建议的重点是确定参与这种选择的蛋白质和机制。研究提出，以了解如何处理双站休息影响这样的决定。我们将使用C。elegans，一种多细胞动物模型系统，使我们能够以低成本和有效的时间尺度在整个生物体中进行DNA修复的遗传分析。利用一个新发现的关键修复基因的突变体，我们将研究在C。秀丽隐杆线虫生殖系此外，我们将确定直接调节参与此过程的关键蛋白质的新机制。为了补充这一分析，我们将通过设计一个系统，针对DNA损伤的特定区域的种系作为一个模型的DNA加工的细胞周期控制进行研究。这种动物模型和人类之间的基因和途径的保守性将成为发现人类中影响癌症形成和发展的基因的有效方法。了解如何选择准确与易出错的通路对于开发利用DNA损伤破坏癌细胞而不伤害健康非癌细胞的抗癌疗法至关重要。