期刊
NUCLEIC ACIDS RESEARCH
卷 49, 期 21, 页码 12048-12068出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkab1124
关键词
-
资金
- National Key Research and Development Program of China [2018YFA0903200]
- National Natural Science Foundation of China
This study identified an enzyme, Bsu06560, capable of metabolizing N-6-methyladenosine and developed mutants F91L and Q150W with enhanced conversion efficiency. Using CRISPR-Cas9, knockout and mutant strains were generated, showing increased N-6-methyladenosine deaminase activity. Transcriptional profiling and gene expression analysis provided insights into the roles of Bsu06560 in regulating adenosine and N-6-methyladenosine metabolism.
N-6-methyladenosine (m(6)A) 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 N-6-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 N-6-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 N-6-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 N-6-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 N-6-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.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据