基础医学与临床 ›› 2023, Vol. 43 ›› Issue (5): 739-744.doi: 10.16352/j.issn.1001-6325.2023.05.0739

• 研究论文 • 上一篇    下一篇

K562细胞中N-乙酰基转移酶10的RNA结合图谱分析

李高原, 王妍然, 王芳, 余佳*   

  1. 中国医学科学院基础医学研究所 北京协和医学院基础学院 医学分子生物学国家重点实验室,北京 100005
  • 收稿日期:2023-02-21 修回日期:2023-03-21 出版日期:2023-05-05 发布日期:2023-04-26
  • 通讯作者: *j-yu@ibms.pumc.edu.cn
  • 基金资助:
    国家杰出青年科学基金(31725013)

RNA binding profile analysis of N-acetyltransferase 10 in K562 cells

LI Gaoyuan, WANG Yanran, WANG Fang, YU Jia*   

  1. State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC, Beijing 100005, China
  • Received:2023-02-21 Revised:2023-03-21 Online:2023-05-05 Published:2023-04-26
  • Contact: *j-yu@ibms.pumc.edu.cn

摘要: 目的 对N-乙酰基转移酶10 (NAT10)在人慢性髓原白血病细胞系K562中的RNA结合图谱进行描绘。方法 利用紫外交联免疫沉淀技术(eCLIP)捕获NAT10在K562细胞中结合的转录本集合,对其进行高通量测序,并对数据进行生物信息学分析。通过peak-calling分析和对peaks进行注释,鉴定NAT10结合位点。并对所结合基因的类型和结合区域进行分析。进一步通过基因功能富集分析,探索其结合的RNA的功能。结果 NAT10主要结合于蛋白质编码基因的转录本,并在3′非翻译区域(3′UTR)富集,预测显示其结合的转录本与DNA损伤修复等功能相关。结论 NAT10可能通过与DNA损伤修复相关信使RNA(mRNA)的3′UTR区域结合,实现基因表达调控作用。

关键词: N-乙酰基转移酶10, K562细胞, 紫外交联免疫沉淀测序技术(eCLIP-seq)

Abstract: Objective To delineate RNA binding profile of NAT10(N-acetyltransferase 10) in human chronic myelogenous leukemia cell line K562. Methods Enhanced UV crosslinking and immunoprecipitation (eCLIP) were used to capture the transcript collection bound by NAT10 in K562 cells. Data were obtained through high throughput sequencing and then analyzed with bioinformatics. The type and regions of NAT10 binding genes were identified via peak-calling and annotation. The function of the binding RNA was further explored by gene function enrichment analysis. Results NAT10 binding transcripts which were enriched in 3′UTR region, were mainly protein coding genes. Further analysis showed that they were functionally associated with DNA damage and repair. Conclusions NAT10 may regulate gene expression by binding to the 3′UTR region of mRNA associated with DNA damage repair.

Key words: N-acetyltransferase 10, K562 cells, enhanced UV crosslinking, immunoprecipitation, and high-throughput sequencing(eCLIP-seq)

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