RECQ5-dependent SUMO2 conjugation of PCNA in the resolution of transcription-replication conflicts

PCNA 的 RECQ5 依赖性 SUMO2 缀合解决转录复制冲突

• 批准号: 10558750
• 负责人: Yilun Liu
• 金额: $ 38.78万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10558750
• 关键词: Biological Assay; Biology; Bypass; Cell Death; Cells; Chromatin; Chromosome Fragile Sites; Communication; Complex; Conflict (Psychology); DNA; DNA Damage; DNA Double Strand Break; DNA Sequence Rearrangement; DNA biosynthesis; DNA lesion; DNA replication fork; DNA-Directed DNA Polymerase; Data; Defect; Deposition; Event; Fractionation; Frequencies; Gene Deletion; Genes; Genetic Transcription; Genomic Instability; Goals; Hematopoietic Neoplasms; Histones; Human; In Vitro; Knockout Mice; Link; Lysine; Malignant Neoplasms; Mediating; Metabolic; Modeling; Modification; Molecular; Molecular Chaperones; Neoplastic Cell Transformation; Pathogenesis; Proliferating Cell Nuclear Antigen; Proteins; Proteomics; Publishing; RECQL5 gene; RNA; RNA Polymerase II; Resolution; Risk Factors; Role; S phase; SUMO1 gene; Site; Solid Neoplasm; Source; Sumoylation Pathway; Transcriptional Regulation; Tumor Suppressor Proteins; Work; Xenograft procedure; cancer cell; cancer risk; cancer type; cell growth; cell transformation; conflict resolution; helicase; homologous recombination; in vitro Assay; innovation; member; mouse model; novel; prevent; recruit; response; transcriptome; ubiquitin-protein ligase; Biological Assay; Biology; Bypass; Cell Death; Cells; Chromatin; Chromosome Fragile Sites; Communication; Complex; Conflict (Psychology); DNA; DNA Damage; DNA Double Strand Break; DNA Sequence Rearrangement; DNA biosynthesis; DNA lesion; DNA replication fork; DNA-Directed DNA Polymerase; Data; Defect; Deposition; Event; Fractionation; Frequencies; Gene Deletion; Genes; Genetic Transcription; Genomic Instability; Goals; Hematopoietic Neoplasms; Histones; Human; In Vitro; Knockout Mice; Link; Lysine; Malignant Neoplasms; Mediating; Metabolic; Modeling; Modification; Molecular; Molecular Chaperones; Neoplastic Cell Transformation; Pathogenesis; Proliferating Cell Nuclear Antigen; Proteins; Proteomics; Publishing; RECQL5 gene; RNA; RNA Polymerase II; Resolution; Risk Factors; Role; S phase; SUMO1 gene; Site; Solid Neoplasm; Source; Sumoylation Pathway; Transcriptional Regulation; Tumor Suppressor Proteins; Work; Xenograft procedure; cancer cell; cancer risk; cancer type; cell growth; cell transformation; conflict resolution; helicase; homologous recombination; in vitro Assay; innovation; member; mouse model; novel; prevent; recruit; response; transcriptome; ubiquitin-protein ligase

Project Summary Proliferating cell nuclear antigen (PCNA) is an essential component of the replisome, and it enhances the processivity of the DNA polymerases in DNA synthesis. In addition, in response to DNA damage, PCNA is ubiquitinated at lysine 164 (K164) to bypass DNA lesions. In unperturbed cells, the same K164 residue can also be conjugated with either SUMO1 or SUMO2, and SUMO1-PCNA has been implicated in recruiting PARI helicase to suppress homologous recombination. However, the source of replication obstacle that triggers PCNA SUMOylation is yet to be defined, and the regulators of PCNA SUMOylation are not known. It is also not clear if SUMO2-PCNA functions redundantly to SUMO1-PCNA. Our newly published data argue that human SUMO2-PCNA has a unique function in transcription that is not shared by SUMO1-PCNA, because only SUMO2-PCNA is associated with transcriptionally active chromatin. Even though PCNA SUMO2 conjugation occurs in S-phase, SUMO2-PCNA is induced by RNA polymerase II - dependent transcription and requires the RNA polymerase II - interacting protein, RECQ5 DNA helicase. Importantly, cells with reduced SUMO2-PCNA accumulate transcription-induced DNA double-strand breaks during S-phase. Therefore, our data support a conceptually innovative model for a role of SUMO2-PCNA in resolving transcription-replication conflicts to minimize genomic instability. The goal of this proposal is (1) to identify the molecular factors that facilitate the transcription-induced SUMO2 conjugation of PCNA, (2) to identify the molecular mechanism by which SUMO2- PCNA resolves transcription-replication conflicts, and (3) to elucidate the contribution of SUMO2-PCNA toward preventing genomic instability and neoplastic transformation, as transcription-replication conflicts are major source for common fragile site instability.

项目摘要 增殖细胞核抗原（PCNA）是重新组合的重要组成部分，它增强了 DNA合成中DNA聚合酶的加工性。另外，针对DNA损伤，PCNA为 在赖氨酸164（K164）中泛素化以绕过DNA病变。在不受干扰的细胞中，相同的K164残基可以 也与SUMO1或SUMO2共轭，SUMO1-PCNA与招募Pari有关 解旋酶抑制同源重组。但是，触发复制障碍物的来源 PCNA sumoylation尚未定义，并且PCNA sumoylation的调节剂尚不清楚。也不 清除SUMO2-PCNA是否可以冗余到SUMO1-PCNA。我们新发布的数据认为人类 SUMO2-PCNA在转录中具有独特的功能，而Sumo1-PCNA并未共享，因为仅 SUMO2-PCNA与转录活性染色质有关。即使PCNA SUMO2共轭 发生在S期，SUMO2-PCNA由RNA聚合酶II-依赖性转录诱导，需要 RNA聚合酶II-相互作用蛋白，RECQ5 DNA解旋酶。重要的是，SUMO2-PCNA还原的细胞 在S期间积累转录诱导的DNA双链断裂。因此，我们的数据支持 概念上创新的模型，用于SUMO2-PCNA在解决转录复制冲突中的作用 最小化基因组不稳定性。该提案的目的是（1）确定促进该分子因素 转录诱导的PCNA的SUMO2共轭，（2），以识别SUMO2-的分子机制 PCNA解决转录复制冲突，（3）阐明Sumo2-PCNA对 防止基因组不稳定和肿瘤转化，因为转录复制冲突是主要的 常见的脆弱现场不稳定性的来源。