DNA-Protein Crosslinks and Genomic Instability

DNA-蛋白质交联和基因组不稳定性

基本信息

批准号：
10533787
负责人：
Matthew J Schellenberg
金额：
$ 35.64万
依托单位：
MAYO CLINIC ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-01-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10533787
关键词：
Active Sites Antibodies Binding Bioinformatics Cancer Etiology Cell Cycle Cell Line Cells Chromosome abnormality Complex DNA DNA Damage DNA Maintenance DNA Repair Gene DNA biosynthesis DNA metabolism DNA replication fork DNA-protein crosslink Data Defect Environmental Carcinogens Enzymes Exhibits Formaldehyde G2 Phase Gene Expression Genes Genetic Genetic Transcription Genome Stability Genomic DNA Genomic Instability Histones Human In Vitro Knock-out Knockout Mice Left Ligation Maintenance Malignant Neoplasms Malignant neoplasm of liver Metalloproteases Modeling Mus Mutation Nature Neuroblastoma Nuclear Peptide Hydrolases Peptides Phenotype Play Point Mutation Predisposition Process Progeria Prognosis Protease Domain Protein Databases Proteins Publishing Reaction Repair Complex Reporting Role Rotation Source Stress Structure Syndrome Testing Therapeutic Trypsin Tumor Suppression Type I DNA Topoisomerases Ubiquitin Zinc Fingers adduct cancer therapy chemotherapy crosslink demethylation ds-DNA experimental study genome integrity insight micronucleus mouse model mutant neoplastic cell neuroblastoma cell novel prevent recruit repaired tumorigenesis

项目摘要

Project Summary/Abstract DNA damage is the major cause of genomic instability, a hallmark of cancer. DNA-protein crosslinks (DPCs) are one of most toxic forms of DNA damage, interfering with the DNA replication and transcription machineries due to their bulky nature. However, despite the importance of repairing DPCs, very little is known about the DPC repair process. Recent studies, including studies from our lab, identified Spartan (SPRTN, also known as DVC1) as a nuclear metalloprotease that associates with PCNA and plays a critical role in DPC repair at DNA replication forks. Our published studies demonstrated that SPRTN hypomorphic mice exhibited genomic instability, progeria, and liver cancer: all of the major phenotypes in human Ruijs-Aalfs Syndrome, which is caused by inactivating mutations in SPRTN. These results underscore the significance SPRTN in DPC repair and the maintenance of genomic stability. However, it is not known whether other proteases are also involved in DPC repair. Here, we describe our preliminary studies of a novel human protease, which we identified using bioinformatics analyses to search for additional human proteins with domain features similar to SPRTN. Like SPRTN, this protease contains a putative protease domain (trypsin-like peptidase) and a PCNA-interacting peptide (PIP) box. Our preliminary experiments showed that depletion of the protease resulted in accumulation of DPCs, leading us to hypothesize that it also participates in DPC repair. Moreover, low levels of expression of the protease in neuroblastoma are associated with increased genomic instability, suggesting that its deficiency causes genomic instability and cancer. To test these hypotheses, we propose to: 1) study the relationship between the novel protease and SPRTN in DPC repair and sensitivity to DPC-inducing chemotherapeutics; 2) define the functional domains required for DPC repair; and 3) examine genome stability and cancer susceptibility in a knockout mouse model. This project will not only elucidate the mechanism of protease- dependent DPC repair, but will also provide new insight into genome instability and tumorigenesis caused by DPCs, and possibly identify new strategies for cancer therapy.

项目摘要/摘要 DNA损伤是基因组不稳定性的主要原因，这是癌症的标志。 DNA-蛋白交联（DPC）是最有毒的DNA损伤形式之一，干扰了DNA复制和转录机器由于他们的本性。但是，尽管维修DPC很重要，但对 DPC维修过程。最近的研究，包括来自我们实验室的研究，确定了Spartan（SPRTN，也称为 DVC1）作为与PCNA相关的核金属蛋白酶，在DNA的DPC修复中起着至关重要的作用复制叉。我们发表的研究表明，SPRTN型肌肉小鼠表现出基因组不稳定性，后代和肝癌：人Ruijs-aalfs综合征的所有主要表型，由SPRTN中的突变灭活引起。这些结果强调了DPC修复中的显着性SPRTN 以及基因组稳定性的维持。但是，尚不知道其他蛋白酶是否也涉及在DPC维修中。在这里，我们描述了我们对新型人类蛋白酶的初步研究，我们使用该研究确定生物信息学分析以搜索具有类似于SPRT的域特征的其他人类蛋白质。喜欢 SPRTN，该蛋白酶包含假定的蛋白酶结构域（胰蛋白酶样肽酶）和PCNA相互作用肽（PIP）盒。我们的初步实验表明，蛋白酶的耗竭导致积累 DPC，导致我们假设它也参与DPC维修。此外，低水平的表达神经母细胞瘤中的蛋白酶与基因组不稳定性增加有关，表明其缺乏症引起基因组不稳定性和癌症。为了检验这些假设，我们建议：1）研究关系在DPC修复中的新型蛋白酶和SPRN之间以及对DPC诱导化学疗法的敏感性之间； 2）定义DPC维修所需的功能域； 3）检查基因组稳定和癌症敲除鼠标模型中的敏感性。该项目不仅将阐明蛋白酶的机制依赖的DPC修复，但也将提供有关基因组不稳定性和肿瘤发生的新见解。 DPC，并可能确定癌症治疗的新策略。