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: 10641022
• 负责人: Gary M. Kupfer
• 金额: $ 48.52万
• 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/10641022
• 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; Co-Immunoprecipitations; Collaborations; Complex; 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; 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; Productivity; Property; Proteins; Publishing; RNA; RNA Splicing; Repair Enzymology; Research Personnel; Resolution; Role; SF1; Scientist; Single-Stranded DNA; Site; Stress; Structure; Structure-Activity Relationship; Substrate Specificity; System; Testing; Time; Transcript; Tumor Suppression; Tumor Suppressor Proteins; 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.

项目概要 基因组完整性的维持取决于细胞修复受损 DNA 和解决问题的能力 转录复制冲突。在这方面，R-环、三链核酸结构 含有与 DNA 模板杂交的 RNA 转录物，可能会损害多种基因组的稳定性 方式。具体来说，R 环结构内的 ssDNA 容易受到溶核切割，从而导致 转录相关诱变或转录相关重组。而且，碰撞 带有 R 环的 DNA 复制机制的破坏会导致复制叉崩溃、DNA 双链 断裂（DSB）、叉融合和染色体易位，这些都会导致肿瘤 转化和肿瘤发生。 我们 MPI 资助的这一竞争性延续充分利用了我们在 DNA 修复酶学方面的独特专业知识 和细胞生物学模型来描绘 R 环解析机制的结构功能 SF1 家族解旋酶 Senataxin (SETX) 和肿瘤抑制复合物 BRCA1-BARD1。在 具体目标1，我们将定义SETX异常通用的核酸解旋活性并进行测试 假设 SETX 通过特定的展开活动直接解决 R 循环。具体目标2 将确定 BRCA1-BARD1 在 SETX 介导的 R 环解析中的作用，以检验以下假设： BRCA1-BARD1 与 SETX 合作，通过询问 SETX 和 我们重建的生化系统和细胞中的 BRCA1-BARD1。 这次 MPI 更新是基于与 Patrick Sung 博士（ 领先的 DNA 修复酶学家和 Gary Kupfer 博士，一位利用遗传技术的医学科学家 Fanconi 贫血模型可促进对 DNA 修复途径和机制的理解。 连同众多具有重要意义的合着论文，我们持续的合作努力 承诺发挥最高程度的影响并提供对机制基础的洞察 与肿瘤抑制途径相关的主要基因组维持途径。

项目成果

Structural insights into 5' flap DNA unwinding and incision by the human FAN1 dimer.

对人类 FAN1 二聚体 5 瓣 DNA 解旋和切割的结构见解。

• 发表时间: 2014
• 影响因子: 16.6
• 作者: Zhao, Qi;Xue, Xiaoyu;Longerich, Simonne;Sung, Patrick;Xiong, Yong
• 通讯作者: Xiong, Yong

A novel role for non-ubiquitinated FANCD2 in response to hydroxyurea-induced DNA damage.

非泛素化 FANCD2 在响应羟基脲诱导的 DNA 损伤中的新作用。

• 发表时间: 2016
• 影响因子: 8
• 作者: Chen, X;Bosques, L;Sung, P;Kupfer, G M
• 通讯作者: Kupfer, G M

Homologous Recombination Repair in Biliary Tract Cancers: A Prime Target for PARP Inhibition?

胆道癌中的同源重组修复：PARP 抑制的主要目标？

• DOI: 10.3390/cancers14102561
• 发表时间: 2022-05-23
• 期刊: Cancers
• 影响因子: 5.2
• 作者: Yin, Chao;Kulasekaran, Monika;Roy, Tina;Decker, Brennan;Alexander, Sonja;Margolis, Mathew;Jha, Reena C.;Kupfer, Gary M.;He, Aiwu R.
• 通讯作者: He, Aiwu R.

Stress and DNA repair biology of the Fanconi anemia pathway.

Fanconi 贫血途径的应激和 DNA 修复生物学。

• DOI: 10.1182/blood-2014-04-526293
• 发表时间: 2014-10-30
• 期刊: Blood
• 影响因子: 20.3
• 作者: S. Longerich;Jian Li;Y. Xiong;P. Sung;G. Kupfer
• 通讯作者: G. Kupfer

Role of RAD51AP1 in homologous recombination DNA repair and carcinogenesis.

RAD51AP1 在同源重组 DNA 修复和癌发生中的作用。

• DOI: 10.1016/j.dnarep.2017.09.008
• 发表时间: 2017-11
• 期刊: DNA repair
• 影响因子: 3.8
• 作者: Pires E;Sung P;Wiese C
• 通讯作者: Wiese C