Investigation of a Novel Role for RNA Binding Proteins in DNA Repair

RNA 结合蛋白在 DNA 修复中的新作用的研究

• 批准号: 8257982
• 负责人: William S. Dynan
• 金额: $ 6.79万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-26 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8257982
• 关键词: Adenosine; Area; Biogenesis; Biology; Cells; Cellular biology; Chromosome Pairing; Clinical; Complex; Core Protein; DNA; DNA Damage; DNA Double Strand Break; DNA Repair; DNA Repair Pathway; Deposition; Distant; Double Strand Break Repair; Family; Family member; Functional RNA; Gene Expression; Gene Expression Regulation; Human; Inosine; Investigation; Ionizing radiation; Lesion; Link; Mammalian Cell; Mediating; Mediator of activation protein; Messenger RNA; Modeling; Molecular; Nature; Normal tissue morphology; Nuclear Structure; Nucleic Acids; Nucleoplasm; Pathway interactions; Phenotype; Post-Translational Protein Processing; Protein Family; Proteins; RNA; RNA-Binding Proteins; Radiation; Radiation Tolerance; Radiation therapy; Radiation-Sensitizing Agents; Relative (related person); Reliance; Research; Role; Site; Structure; System; Testing; Therapeutic; Untranslated RNA; Work; genetic manipulation; homologous recombination; improved; innovation; interest; member; novel; protein function; public health relevance; radiation resistance; reconstitution; repaired; research study; response; scaffold; tumor

DESCRIPTION (provided by applicant): This is a proposal to investigate proteins that contribute to the efficiency and accuracy of DNA double-strand break (DSB) repair. Ionizing radiation deposits energy along discrete tracks, resulting in clustered DNA damage and DSBs. The ability to repair these signature lesions is a major determinant of radiation sensitivity and resistance in both normal tissue and tumors. Manipulation of DNA repair pathways therefore affords a promising approach for improving the efficacy of radiotherapy. Recent work provides evidence for the novel involvement of a small family of human RNA binding proteins in DSB repair. These proteins are core components of paraspeckles, which are nuclear structures that are organized around a long noncoding RNA scaffold and that regulate gene expression by retaining adenosine-to- inosine hyper-edited mRNAs. Separate experiments indicate, however, that these proteins participate in both homologous recombination and nonhomologous end joining, which are the two main pathways of DSB repair in human cells. The three members of the family in humans-PSF, p54nrb, and PSPC1-rapidly relocalize to sites of induced DNA damage, suggesting the existence of a molecular switch that controls RNA versus DNA interaction. The hypothesis to be tested is that PSF and its partners are mediators of gene regulation and DNA repair that switch rapidly between RNA and DNA interaction modes following the induction of DNA damage. A unifying theme may be reliance on an intrinsic ability of PSF and its partners to promote pairing of distant nucleic acid segments. The first specific aim will be to test a prediction that a PSF7p54nrb complex promotes juxtaposition of opposing DNA ends in a loop structure. The second will be to examine repair functions of PSF and its partners more broadly using genetic manipulation of human cells. The third will focus directly on how PSF and its partners switch between RNA biogenesis and DSB repair modes and will investigate the therapeutic applicability of this mechanism. The proposed research is innovative, because the primary sequence and domain structure of PSF, p54nrb, and PSPC1 are unlike any previously characterized DSB repair proteins. The work is scientifically significant, because it explores a previously unsuspected link between DSB repair and the biology of non-coding RNAs, which is an interesting and topical area in cell biology. Finally, the work has potential clinical and translational impact, because of the possibility that therapeutic RNAs might be developed to influence switching between RNA biogenesis and DNA repair modes to alter clinical radiation response. PUBLIC HEALTH RELEVANCE: This is a proposal to investigate the novel role of a family of RNA binding proteins DNA double-strand break (DSB) repair. The ability to repair DSBs is a major determinant of radiation sensitivity and resistance in both normal tissue and tumors. The project has potential clinical and translational impact because of the possibility that therapeutic RNAs might be developed to inhibit the DNA repair activity of these proteins and thus alter the clinical radiation response.

描述（由申请人提供）：这是一项研究有助于 DNA 双链断裂 (DSB) 修复效率和准确性的蛋白质的提案。电离辐射沿着离散轨迹沉积能量，导致聚集的 DNA 损伤和 DSB。修复这些标志性病变的能力是正常组织和肿瘤的辐射敏感性和抵抗力的主要决定因素。因此，DNA 修复途径的调控为提高放射治疗的疗效提供了一种有前景的方法。最近的工作为人类 RNA 结合蛋白小家族新参与 DSB 修复提供了证据。这些蛋白质是副斑点的核心成分，副斑点是围绕长非编码 RNA 支架组织的核结构，通过保留腺苷到肌苷超编辑的 mRNA 来调节基因表达。然而，单独的实验表明，这些蛋白质参与同源重组和非同源末端连接，这是人类细胞中 DSB 修复的两个主要途径。人类中该家族的三个成员 - PSF、p54nrb 和 PSPC1 - 迅速重新定位到诱导 DNA 损伤的位点，表明存在控制 RNA 与 DNA 相互作用的分子开关。待检验的假设是，PSF 及其伙伴是基因调控和 DNA 修复的介导者，在诱导 DNA 损伤后，它们在 RNA 和 DNA 相互作用模式之间快速切换。一个统一的主题可能是依赖 PSF 及其伙伴促进远距离核酸片段配对的内在能力。第一个具体目标是测试 PSF7p54nrb 复合物促进环结构中相反 DNA 末端并置的预测。第二个目标是利用人类细胞的基因操作来更广泛地检查 PSF 及其伙伴的修复功能。第三部分将直接关注 PSF 及其合作伙伴如何在 RNA 生物发生和 DSB 修复模式之间切换，并将研究该机制的治疗适用性。拟议的研究具有创新性，因为 PSF、p54nrb 和 PSPC1 的一级序列和结构域与任何先前表征的 DSB 修复蛋白不同。这项工作具有重要的科学意义，因为它探索了 DSB 修复与非编码 RNA 生物学之间以前未被怀疑的联系，这是细胞生物学中一个有趣且热门的领域。最后，这项工作具有潜在的临床和转化影响，因为治疗性 RNA 可能被开发来影响 RNA 生物发生和 DNA 修复模式之间的切换，从而改变临床放射反应。 公共健康相关性：这是一项研究 RNA 结合蛋白家族 DNA 双链断裂 (DSB) 修复的新作用的提案。修复 DSB 的能力是正常组织和肿瘤的辐射敏感性和抵抗力的主要决定因素。该项目具有潜在的临床和转化影响，因为治疗性 RNA 可能被开发来抑制这些蛋白质的 DNA 修复活性，从而改变临床放射反应。