Mechanistic Dissection of the BRCA1-SETX-dependent Pathway of R-loop Avoidance and Genome Maintenance

R 环避免和基因组维护的 BRCA1-SETX 依赖性途径的机制剖析

• 批准号: 10537108
• 负责人: Gary M. Kupfer
• 金额: $ 50.88万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10537108
• 关键词: Adult; Advanced Malignant Neoplasm; BARD1 gene; BRCA1 gene; Biochemical; Biological; Biological Assay; Biological Process; Biology; Bone marrow failure; Cell Extracts; Cell Survival; Cell physiology; Cells; Cellular biology; Chromosomal translocation; Collaborations; Complex; Conflict (Psychology); Coupled; DNA; DNA Damage; DNA Double Strand Break; DNA Repair; DNA Repair Pathway; DNA Structure; DNA biosynthesis; DNA lesion; DNA replication fork; Data; Diagnosis; Disease; Dissection; Event; Family; Fanconi' s Anemia; Genetic Models; Genetic Recombination; Genetic Transcription; Genome; Genome Stability; Grant; Head; Hematopoiesis; Hybrids; In Vitro; Joint Ventures; Lead; Light; Link; Maintenance; Malignant Childhood Neoplasm; Malignant Neoplasms; Maps; Mediating; Messenger RNA; Methods; Modeling; Mutagenesis; Mutation; Nature; Neoplastic Cell Transformation; Nucleic Acid Amplification Tests; Nucleic Acid Binding; Nucleic Acids; Obstruction; Paper; Pathologic; Pathway interactions; Pediatric Oncologist; Physicians; Physiological; Process; Property; Proteins; Publishing; RNA; RNA Splicing; Repair Enzymology; Research Personnel; Resolution; Role; SF1; Scientist; Site; Stress; Structure; Structure-Activity Relationship; Substrate Specificity; System; Testing; Time; Transcript; Tumor Suppression; Tumor Suppressor Proteins; base; cancer prevention; causal variant; genome integrity; helicase; insight; multidisciplinary; mutant; nervous system disorder; novel; nucleic acid structure; preservation; reconstitution; replication stress; response; synergism; tumorigenesis

Project Summary Maintenance of genomic integrity depends on the ability of cells to repair damaged DNA and resolve transcription-replication conflicts. In this regard, R-loops, three-stranded nucleic acid structures that harbor an RNA transcript hybridized to a DNA template, can compromise genome stability in multiple ways. Specifically, the ssDNA within the R-loop structure is vulnerable to nucleolytic cleavage, resulting in transcription-associated mutagenesis or transcription-associated recombination. Moreover, collisions of the DNA replication machinery with R-loops can cause replication fork collapse, DNA double-strand breaks (DSBs), fork fusions, and chromosome translocations, which can then lead to neoplastic transformation and tumorigenesis. This competitive continuation of our MPI grant leverages our unique expertise in DNA repair enzymology and cell biology modeling to delineate the structure-function of an R-loop resolution machinery comprised of the SF1 family helicase Senataxin (SETX) and the tumor suppressor complex BRCA1-BARD1. In Specific Aim 1, we will define the unusually versatile nucleic acid unwinding activity of SETX and test the hypothesis that SETX resolves R loops directly through specific unwinding activity. Specific Aim 2 will determine the role of BRCA1-BARD1 in SETX-mediated R-loop resolution to test the hypothesis that BRCA1-BARD1 cooperates with SETX to resolve pathological R-loops by interrogating SETX and BRCA1-BARD1 in our reconstituted biochemical systems and in cells. This MPI renewal is based on the longstanding and productive collaboration between Dr Patrick Sung, a leading DNA repair enzymologist, and Dr Gary Kupfer, a physician-scientist who has utilized the genetic model of Fanconi anemia to advance understanding of DNA repair pathways and mechanisms. Together, with numerous coauthored papers of high significance, our continuing collaborative endeavors promise to exert impact of the highest degree and to provide insight into the mechanistic underpinnings of a major genome maintenance pathway that is linked to tumor suppression pathways.

项目摘要 基因组完整性的维持取决于细胞修复受损DNA并解决的能力 转录复制冲突。在这方面，R环，三链核酸结构 携带与DNA模板杂交的RNA转录本可以损害多个的基因组稳定性 方式。具体而言，R环结构内的ssDNA容易受到核酸裂解的影响，导致 在转录相关的诱变或转录相关的重组中。而且，碰撞 带有R环的DNA复制机制可能导致复制叉塌陷，DNA双链 断裂（DSB），叉融合和染色体易位，然后会导致肿瘤 转化和肿瘤发生。 MPI赠款的这种竞争延续利用了我们在DNA维修酶学方面的独特专业知识 和细胞生物学建模，以描绘R-Loop分辨率机械的结构功能包括 SF1家族解旋酶塞纳汀（SETX）和肿瘤抑制剂复合物BRCA1-BARD1在 特定的目标1，我们将定义SETX和测试的异常用途的核酸放松活性 SETX通过特定的放松活动直接解决R循环的假设。具体目标2 将确定BRCA1-BARD1在SETX介导的R-LOOP分辨率中的作用，以检验以下假设。 BRCA1-BARD1与SETX合作，通过询问SETX和 在我们重组的生化系统和细胞中的BRCA1-BARD1。 此MPI更新基于帕特里克·昂格博士之间的长期合作， 领先的DNA修复酶学学家和使用遗传学的医师科医师Gary Kupfer博士 Fanconi贫血模型，以提高对DNA修复途径和机制的了解。 我们持续的合作努力共同撰写了许多具有很高意义的合作论文 承诺发挥最高程度的影响，并洞悉机械基础 与肿瘤抑制途径相关的主要基因组维持途径。