Mechanisms of Response to DNA Damage Nuclear Factors

DNA 损伤核因素的反应机制

• 批准号: 7618747
• 负责人: FRIDA E KLEIMAN
• 金额: $ 35.66万
• 依托单位: HUNTER COLLEGE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7618747
• 关键词: Accounting; Applications Grants; BARD1 gene; BRCA1 gene; Biochemical; Biological; Biological Assay; Cells; Cockayne Syndrome; Complex; DNA Damage; DNA Repair; Data; Defect; Development; Disease; Event; Gene Expression; Genetic Transcription; Goals; Mediating; Messenger RNA; Modeling; Nuclear; Paper; Pathway interactions; Phosphorylation; Physiological; Play; Poly(A) Tail; Poly(A)-specific ribonuclease; Polyadenylation; Pre-mRNA Polyadenylation Factor; Process; Productivity; Progress Reports; Proteins; Publications; RNA Polymerase II; RNA Processing; Reaction; Recovery; Regulation; Research; Research Project Grants; Role; TP53 gene; Testing; Transcription-Coupled Repair; Tumor Suppression; Tumor Suppressor Proteins; UV induced; Ubiquitination; United States National Institutes of Health; Work; base; carcinogenesis; cell growth; follow-up; innovation; mRNA Precursor; mRNA Stability; multicatalytic endopeptidase complex; repaired; response; ultraviolet irradiation

DESCRIPTION (provided by applicant): Following UV irradiation the cellular mRNA levels decrease, reflecting a coordinated interaction of DNA repair, transcription, and RNA processing factors. The poly(A) tail is important in the regulation of mRNA turnover and it is fundamental for the control of gene expression. The long-range goal of this research project is to better understand the basic mechanisms of RNA processing regulation upon DNA damage conditions and its physiological significance. The specific hypothesis is that the polyadenylation factor CstF-50 plays an important role in coordinating this nuclear response. We base that hypothesis on these observations: (1) Polyadenylation is inhibited after DNA damage as a result of the formation of the BRCA1/ BARD1/ CstF inhibitory complex and of the proteasome-mediated degradation of the polyadenylation activator RNA polymerase II (RNAP II); (2) DNA damage-induced BARD1 phosphorylation is critical for the UV-induced inhibition of polyadenylation and of RNAP II degradation; (3) CstF functions in the transcription-coupled repair response; (4) CstF-50 can interact with poly(A)-specific ribonuclease (PARN) and regulate its deadenylation activity. Based on these observations and preliminary data, the specific aims are to: (1) Characterize the function of the RNA processing factor CstF in TCR. The role of CstF in different events of the TCR response will be analyzed. Cockayne's syndrome (CS) cells, which are deficient in TCR, will be used. The direct role of CstF and other factors in UV-induced ubiquitination of RNAP II by BRCA1/ BARD1 and in the repair process will also be determined using already proven biochemical assays. (2) Determine the biological significance of the interaction of CstF-50 with PARN. We will determine the effect of these interacting factors on polyadenylation complex formation on the polyadenylation reaction and on the recovery of cellular mRNA levels after DNA damage. Proven biochemical assays and cells deficient in these proteins will be used. (3) Developmental goals: increase the productivity of the research work, which will allow the submission of new papers for publication and the follow-up for an R01 grant application to NIH. The proposed work is innovative because it reflects a functional-mechanistic overlapping of tumor suppressors, like BRCA1/BARD1, and the ubiquitous gene expression machinery, and it suggests a central role for an RNA processing factor in the intricate DNA damage response.

描述（由申请人提供）：紫外线照射后，细胞mRNA水平降低，反映了DNA修复、转录和RNA加工因子的协调相互作用。poly（A）尾在mRNA周转的调节中是重要的，并且是控制基因表达的基础。本研究项目的长期目标是更好地了解RNA加工调节DNA损伤条件的基本机制及其生理意义。具体的假设是，多聚腺苷酸化因子CstF-50在协调这种核反应中起着重要作用。我们的假设基于以下观察结果：（1）DNA损伤后，由于BRCA 1/BARD 1/ CstF抑制复合物的形成和蛋白酶体介导的多聚腺苷酸化激活剂RNA聚合酶II（RNAP II）的降解，多聚腺苷酸化被抑制;（2）DNA损伤诱导的BARD 1磷酸化对于UV诱导的多聚腺苷酸化和RNAP II降解的抑制至关重要;（3）CstF在转录偶联修复反应中发挥作用;（4）CstF-50可与多聚腺苷酸特异性核糖核酸酶（PARN）相互作用并调节其去腺苷化活性。 基于这些观察和初步数据，具体的目的是：（1）表征RNA加工因子CstF在TCR中的功能。将分析CstF在TCR应答的不同事件中的作用。将使用TCR缺陷的科凯恩综合征（CS）细胞。CstF和其他因子在BRCA 1/BARD 1引起的RNAP II的UV诱导泛素化和修复过程中的直接作用也将使用已经证实的生化测定来确定。(2)确定CstF-50与PARN相互作用的生物学意义。我们将确定这些相互作用的因素对多聚腺苷酸化复合物的形成、多聚腺苷酸化反应和DNA损伤后细胞mRNA水平的恢复的影响。将使用非生物化学测定和这些蛋白质缺陷的细胞。(3)发展目标：提高研究工作的生产力，这将允许提交新的论文出版和后续的R 01补助金申请NIH。 这项工作是创新的，因为它反映了肿瘤抑制因子（如BRCA 1/BARD 1）和无处不在的基因表达机制的功能机制重叠，并且它表明RNA加工因子在复杂的DNA损伤反应中起着核心作用。