基础医学与临床 ›› 2021, Vol. 41 ›› Issue (5): 630-635.

• 研究论文 • 上一篇    下一篇

人胚胎肾细胞系HEK293中的翻译调控

柳满, 任悦, 高玉风, 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:2021-01-20 修回日期:2021-03-20 出版日期:2021-05-05 发布日期:2021-05-06
  • 通讯作者: *j-yu@ibms.pumc.edu.cn
  • 基金资助:
    国家自然科学基金(81970103)

Translational regulation of human embryonic kidney cell line HEK293

LIU Man, REN Yue, GAO Yu-feng, 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:2021-01-20 Revised:2021-03-20 Online:2021-05-05 Published:2021-05-06
  • Contact: *j-yu@ibms.pumc.edu.cn

摘要: 目的 探讨人胚胎肾细胞系HEK293中的翻译调控。方法 通过 Polysome profiling结合Western blot实验检测并验证HEK293细胞中mRNA的翻译活性;通过针对RPL10A和RPS17两种核糖体蛋白的RNA免疫共沉淀、建库测序(RIP-seq)及生物信息学分析,获得参与 HEK293细胞蛋白质合成调控的相关分子。结果 在两种核糖体蛋白的RNA免疫共沉淀实验富集的分子中共有129种基因,其中93.8% 为蛋白质编码基因;且功能与RNA剪接、分解代谢、MAPK信号通路、染色体定位、RNA转运等相关。结论 通过针对核糖体蛋白的 RNA 免疫共沉淀可以获得HEK293细胞中活跃翻译的mRNA。

关键词: 人胚胎肾细胞系 293, Polysome profiling, RNA 结合蛋白, 免疫共沉淀

Abstract: Objective To investigate the translational regulation of human embryonic kidney (HEK)293 cell line. Methods Polysome profiling coupled with Western blot was used to verify the translational activity of HEK293 cells. RPL10A and RPS17 RNA immunoprecipitation coupled with sequencing (RIP-seq) and bioinformatic analysis were used to investigate the actively translating mRNAs. Results A total of 129 genes were found in the RNA immunoprecipitation experiments RPL10A and RPS17, among which 93.8% were protein-coding genes and enriched in RNA splicing, metabolism, MAPK signal pathway, chromatin localization and RNA transport. Conclusions A variety of molecules involved in HEK293 cells translation are obtained by RPL10A and RPS17 RIP-seq analysis.

Key words: human embryonic kidney 293 cell line, Polysome profiling, RNA binding protein, immunoprecipitation

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