DNA-guided photoactivatable probe-based chemical proteomics reveals the reader protein of mRNA methylation.

DNA-guided photoactivatable probe-based chemical proteomics reveals the reader protein of mRNA methylation.
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DNA 引导的基于光激活探针的化学蛋白质组学揭示了 mRNA 甲基化的读取蛋白

DOI:
10.1016/j.isci.2021.103046
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发表时间:
2021-09-24
期刊:
影响因子:
5.8
通讯作者:
Zhang K
Zhang K
中科院分区:
综合性期刊2区
文献类型:
--
作者:
Huang Y;Bai X;Guo Z;Dong H;Fu Y;Zhang H;Zhai G;Tian S;Wang Y;Zhang K

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对mRNA的化学修饰可以招募特定的结合蛋白(读取器/伙伴)来确定转录后基因的调控。然而,由于mRNA修饰和阅读器之间的非共价相互作用相当弱和高度动态,阅读器的识别非常有限,因此敏感和稳健的方法是可取的。在这里,我们报告了一种DNA引导的基于光激活的化学蛋白质组学方法,用于分析mRNA甲基化的读取器。以N6-甲基腺苷(M6A)为例,我们证明该方法可以成功地用于标记和丰富mRNA修饰阅读器,以及发现新的合作伙伴。因此,我们将这一策略应用于新的修饰2‘-O-甲基腺苷。结果表明,DDX1是一种潜在的结合蛋白。因此,我们的研究为鉴定mRNA修饰的结合因子提供了一种强有力的化学蛋白质组学工具,并揭示了mRNA修饰的潜在功能。用于共价捕获弱RNA-蛋白质相互作用的DNA模板探针选择性标记和富集甲基化结合因子DDX1被鉴定为2‘-O-甲基腺苷的潜在结合蛋白化学蛋白质组学工具用于确定RNA修饰的结合伙伴生物化学方法;分子生物学;蛋白质组学
Chemical modification on mRNA can recruit specific binding proteins (readers/partners) to determine post-transcriptional gene regulation. However, the identification of the reader is extremely limited owing to the rather weak and highly dynamic non-covalent interactions between mRNA modification and reader, and therefore the sensitive and robust approaches are desirable. Here, we report a DNA-guided photoactivatable-based chemical proteomic approach for profiling the readers of mRNA methylation. By use of N6-methyladenosine (m6A), we illustrated that this method can be successfully utilized for labelling and enriching the readers of mRNA modification, as well as for the discovery of new partners. Thus we applied this strategy to a new modification 2′-O-methyladenosine. As a result, DDX1 was identified and verified as a potential binding protein. Our study therefore provides a powerful chemical proteomics tool for identifying the binding factors of mRNA modification and reveals the underlying function of mRNA modification. A DNA-templated probe for covalent capture of the weak RNA-protein interactions Selective labelling and enrichment of binding factors to mRNA methylation DDX1 was identified as a potential binding protein of 2′-O-methyladenosine A chemical proteomics tool for identifying the binding partners of RNA modifications Biochemistry methods; Molecular biology; Proteomics
n(6) - 甲基腺苷依赖性RNA结构开关调节RNA蛋白质相互作用。
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