拟南芥EBP1蛋白与RNA相互作用的初步研究

doi:10.13560/j.cnki.biotech.bull.1985.2019-1066

摘要/Abstract

摘要： 基因表达调控是现代分子生物学研究的热点话题,随着研究手段的不断成熟,其RNA转录后调控已成为备受关注的领域。RNA结合蛋白（RBPs）是转录后调控的关键因子,参与RNA可变剪切、RNA稳定、翻译等多个过程的调控。ErbB3绑定蛋白（EBP1）是结构功能高度保守的RNA结合蛋白,可与rRNA、tRNA、mRNA等多种RNA结合,参与调控核糖体生物生成、蛋白质翻译多个过程,但目前已报道的靶RNA甚少,对靶RNA的作用机制目前仍不清楚。通过生物信息学分析发现拟南芥EBP1结合RNA的保守结构域,并在体外可直接结合“GUCUCUCACUGCGACGGCUU”序列;通过RNA免疫共沉淀实验找到了3个靶mRNA（AT1G24792、AT3G25211、AT3G24320）;结合核糖体提取及虫草素处理实验发现EBP1可显著调控特定靶mRNA的稳定性及其翻译速率。研究结果不仅证实拟南芥EBP1具有结合RNA的功能,还显示其参与调控靶RNA的转录后事件,为进一步研究EBP1在转录后调控的生物学机制奠定基础。

关键词: EBP1, RNA结合蛋白, mRNA稳定性和翻译

Abstract: Gene expression regulation is a hot spot in modern molecular biology,and the post-transcriptional regulation of RNA has attracted great concerns of researchers with the development of technology. RNA binding proteins（RBPs）,as an important post-transcriptional regulator,are involved in RNA alternative splicing,RNA stability,translation,and so on. ErbB3 binding protein（EBP1）is an RBP with highly conserved structure and functions,and is involved in the regulation of ribosome biogenesis and protein translation through binding a variety of RNAs such as rRNA,tRNA,and mRNA. However,few of RNA targets of EBP1 have been documented and the mechanism of interaction with its RNA target is still unknown. Here,the conserved RNA-binding domain in Arabidopsis EBP1 was predicted through bioinformatics analysis. “GUCUCUCACUGCGACGGCUU” sequence was directly bound by EBP1 in vitro. Further,mRNAs of three genes（AT1G24792,AT3G25211 and AT3G24320）were identified in RNA immunoprecipitation assay. Polysome profiling and cordycepin treatment experiments demonstrated that EBP1 significantly regulated the stability of these target mRNAs and their translation rates. Our results confirm that Arabidopsis EBP1 has the function of binding RNA,and regulates the post-transcriptional events of target RNAs,which provides useful data for further studying the biological mechanism of EBP1 post-transcriptional regulation.

Key words: EBP1, RNA binding protein, mRNA stability and translation

李晓燕, 李驰宇, 于峰, 廖红东. 拟南芥EBP1蛋白与RNA相互作用的初步研究[J]. 生物技术通报, 2020, 36(6): 35-45.

LI Xiao-yan, LI Chi-yu, YU Feng, LIAO Hong-dong. Preliminary Study on Interaction Between Arabidopsis thaliana EBP1 Protein and RNA[J]. Biotechnology Bulletin, 2020, 36(6): 35-45.

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