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DOI	10.1073/pnas.2026183118
	Use of NAD tagSeq II to identify growth phase-dependent alterations in E. coli RNA NAD+ capping
	Zhang H.; Zhong H.; Wang X.; Zhang S.; Shao X.; Hu H.; Yu Z.; Cai Z.; Chen X.; Xia Y.
发表日期	2021
ISSN	00278424
卷号	118 期号:14
英文摘要	Recent findings regarding nicotinamide adenine dinucleotide (NAD+)-capped RNAs (NAD-RNAs) indicate that prokaryotes and eukaryotes employ noncanonical RNA capping to regulate gene expression. Two methods for transcriptome-wide analysis of NADRNAs, NAD captureSeq and NAD tagSeq, are based on coppercatalyzed azide-alkyne cycloaddition (CuAAC) click chemistry to label NAD-RNAs. However, copper ions can fragment/degrade RNA, interfering with the analyses. Here we report development of NAD tagSeq II, which uses copper-free, strain-promoted azide-alkyne cycloaddition (SPAAC) for labeling NAD-RNAs, followed by identification of tagged RNA by single-molecule direct RNA sequencing. We used this method to compare NAD-RNA and total transcript profiles of Escherichia coli cells in the exponential and stationary phases. We identified hundreds of NAD-RNA species in E. coli and revealed genome-wide alterations of NAD-RNA profiles in the different growth phases. Although no or few NAD-RNAs were detected from some of the most highly expressed genes, the transcripts of some genes were found to be primarily NAD-RNAs. Our study suggests that NAD-RNAs play roles in linking nutrient cues with gene regulation in E. coli. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	E. coli; Gene regulation; NAD tagSeq II; NAD+-capped RNAs; SPAAC
语种	英语
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/179991
作者单位	Department of Biology, Hong Kong Baptist University, Hong Kong, Hong Kong; Department of Botany and Plant Sciences, Institute of Integrative Genome Biology, University of California, Riverside, CA 92521, United States; State Key Laboratory of Agrobiotechnology, The Chinese University of Hong Kong, Hong Kong, Hong Kong; Centre for Soybean Research, School of Life Sciences, The Chinese University of Hong Kong, Hong Kong, Hong Kong; State Key Laboratory of Environmental and Biological Analysis, Department of Chemistry, Hong Kong Baptist University, Hong Kong, Hong Kong; School of Chinese Medicine, Hong Kong Baptist University, Hong Kong, Hong Kong
推荐引用方式 GB/T 7714	Zhang H.,Zhong H.,Wang X.,et al. Use of NAD tagSeq II to identify growth phase-dependent alterations in E. coli RNA NAD+ capping[J],2021,118(14).
APA	Zhang H..,Zhong H..,Wang X..,Zhang S..,Shao X..,...&Xia Y..(2021).Use of NAD tagSeq II to identify growth phase-dependent alterations in E. coli RNA NAD+ capping.Proceedings of the National Academy of Sciences of the United States of America,118(14).
MLA	Zhang H.,et al."Use of NAD tagSeq II to identify growth phase-dependent alterations in E. coli RNA NAD+ capping".Proceedings of the National Academy of Sciences of the United States of America 118.14(2021).