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酶促脱氨作用使表观遗传核苷 N6-甲基腺苷调控基因表达。

Enzymatic deamination of the epigenetic nucleoside N6-methyladenosine regulates gene expression.

机构信息

The Institute of Advanced Studies, and Key Laboratory of Biomedical Polymers-Ministry of Education, College of Chemistry and Molecular Sciences, Wuhan University, 40072 Wuhan, P.R. China.

出版信息

Nucleic Acids Res. 2021 Dec 2;49(21):12048-12068. doi: 10.1093/nar/gkab1124.

DOI:10.1093/nar/gkab1124
PMID:34850126
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8643624/
Abstract

N6-methyladenosine (m6A) modification is the most extensively studied epigenetic modification due to its crucial role in regulating an array of biological processes. Herein, Bsu06560, formerly annotated as an adenine deaminase derived from Bacillus subtilis 168, was recognized as the first enzyme capable of metabolizing the epigenetic nucleoside N6-methyladenosine. A model of Bsu06560 was constructed, and several critical residues were putatively identified via mutational screening. Two mutants, F91L and Q150W, provided a superiorly enhanced conversion ratio of adenosine and N6-methyladenosine. The CRISPR-Cas9 system generated Bsu06560-knockout, F91L, and Q150W mutations from the B. subtilis 168 genome. Transcriptional profiling revealed a higher global gene expression level in BS-F91L and BS-Q150W strains with enhanced N6-methyladenosine deaminase activity. The differentially expressed genes were categorized using GO, COG, KEGG and verified through RT-qPCR. This study assessed the crucial roles of Bsu06560 in regulating adenosine and N6-methyladenosine metabolism, which influence a myriad of biological processes. This is the first systematic research to identify and functionally annotate an enzyme capable of metabolizing N6-methyladenosine and highlight its significant roles in regulation of bacterial metabolism. Besides, this study provides a novel method for controlling gene expression through the mutations of critical residues.

摘要

N6-甲基腺苷（m6A）修饰是研究最广泛的表观遗传修饰，因为它在调节一系列生物过程中起着至关重要的作用。在此，Bsu06560 被鉴定为第一个能够代谢表观遗传核苷 N6-甲基腺苷的酶，它以前被注释为来自枯草芽孢杆菌 168 的腺嘌呤脱氨酶。构建了 Bsu06560 的模型，并通过突变筛选推测出几个关键残基。两个突变体 F91L 和 Q150W 提供了更高的腺苷和 N6-甲基腺苷转化率。CRISPR-Cas9 系统从枯草芽孢杆菌 168 基因组中生成了 Bsu06560 敲除、F91L 和 Q150W 突变。转录谱分析显示，具有增强的 N6-甲基腺苷脱氨酶活性的 BS-F91L 和 BS-Q150W 菌株具有更高的全局基因表达水平。使用 GO、COG、KEGG 对差异表达基因进行分类，并通过 RT-qPCR 进行验证。本研究评估了 Bsu06560 在调节腺苷和 N6-甲基腺苷代谢中的关键作用，这些代谢影响着无数的生物过程。这是首次对能够代谢 N6-甲基腺苷的酶进行鉴定和功能注释的系统研究，并强调了其在细菌代谢调控中的重要作用。此外，本研究通过关键残基的突变提供了一种控制基因表达的新方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2626/8643624/271bbc16b9c6/gkab1124fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2626/8643624/049340cf8b36/gkab1124fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2626/8643624/5879cb973e0a/gkab1124fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2626/8643624/80f46ffe7081/gkab1124fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2626/8643624/202813a67a00/gkab1124fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2626/8643624/271bbc16b9c6/gkab1124fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2626/8643624/049340cf8b36/gkab1124fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2626/8643624/5879cb973e0a/gkab1124fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2626/8643624/80f46ffe7081/gkab1124fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2626/8643624/202813a67a00/gkab1124fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2626/8643624/271bbc16b9c6/gkab1124fig5.jpg

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