摘要
目的寻找新的参与DNA复制应激响应的相关蛋白,探究其通过影响DNA复制导致基因组不稳定的机制。方法利用TurboID邻近蛋白标记技术,质谱鉴定DNA复制应激下邻近单链DNA结合蛋白的蛋白质组学,筛选参与基因组稳定相关的新功能蛋白。通过慢病毒介导的目标蛋白敲低以及过表达,利用Western blot及免疫荧光等探索TP53RK蛋白对基因组稳定性的调控作用。结果TP53RK响应DNA复制应激,敲低TP53RK导致细胞增殖速率减慢,基因组不稳定标志蛋白γH2AX明显升高。同时敲低TP53RK后细胞对DNA复制应激诱导剂更为敏感,过表达TP53RK则可以缓解DNA复制应激对细胞造成的损伤。结论通过筛选得到多个与DNA复制应激响应相关的蛋白,其中TP53RK通过影响细胞对DNA复制应激的响应来改变基因组的稳定性,可能与多种相关疾病的发生密切相关。
Objective:To identify the DNA replication stress response-associated proteins and explore their roles in maintaining genome stability during DNA replication.Methods:In this study,we utilized the TurboID-based technique to identify the proteomics adjacent to single-stranded DNA binding proteins under DNA replication stress,and to identify the specific proteins in genomic stability maintenance.Subsequently,we analyzed the genome stability in cells with lentivirusmediated knockdown of target gene via Western blot and immunofluorescence.Results:We found that TP53RK responds to DNA replication stress.Knockdown of TP53RK leads to a slower cell proliferation rate,and a significant increase in the expression of the genomic instability marker proteinγH2AX,which indicates that the cells more sensitive to DNA replication stress.Moreover,overexpression of TP53RK alleviates the damage to cells caused by DNA replication stressinducing agents.Conclusion:TP53RK affects genomic stability by influencing the cellular response to DNA replication stress and may be closely related to the occurrence of multiple diseases.
作者
吕梦
于志远
吕新星
LV Meng;YU Zhiyuan;LV Xinxing(School of Clinical and Basic Medical Sciences,Shandong First Medical University&Shandong Academy of Medical Sciences,Jinan 250117,China;Central Laboratory of Hospital for Skin Diseases,Shandong First Medical University,Jinan 250022,China)
出处
《山东第一医科大学(山东省医学科学院)学报》
2025年第7期412-419,共8页
Journal of Shandong First Medical University & Shandong Academy of Medical Sciences
基金
山东省自然科学基金(ZR2021QC030,ZR2024MC179)。
