N(6)-allyladenosine: A new small molecule for RNA labeling identified by mutation assay Journal Article

Authors: Shu, X.; Dai, Q.; Wu, T.; Bothwell, I. R.; Yue, Y.; Zhang, Z.; Cao, J.; Fei, Q.; Luo, M.; He, C.; Liu, J.
Article Title: N(6)-allyladenosine: A new small molecule for RNA labeling identified by mutation assay
Abstract: RNA labeling is crucial for the study of RNA structure and metabolism. Herein we report N6-allyladenosine (a6A) as a new small molecule for RNA labeling through both metabolic and enzyme-assisted manners. a6A behaves like A and can be metabolically incorporated into newly synthesized RNAs inside mammalian cells. We also show that human RNA N6-methyladenosine (m6A) methyltransferases METTL3/METTL14 can work with a synthetic cofactor, namely allyl-SAM (S-adenosyl methionine with methyl replaced by allyl) in order to site-specifically install an allyl group to the N6-position of A within specific sequence to generate a6A-labeled RNAs. The iodination of N6-allyl group of a6A under mild buffer conditions spontaneously induces the formation of N1,N6-cyclized adenosine and creates mutations at its opposite site during complementary DNA synthesis of reverse transcription. The existing m6A in RNA is inert to methyltransferase-assisted allyl labeling, which offers a chance to differentiate m6A from A at individual RNA sites. Our work demonstrates a new method for RNA labeling, which could find applications in developing sequencing methods for nascent RNAs and RNA modifications. © 2017 American Chemical Society.
Keywords: human cell; dna synthesis; metabolism; rna; methyltransferase; methyltransferases; rna synthesis; nucleic acids; mutation rate; amino acids; cyclization; matrix assisted laser desorption ionization time of flight mass spectrometry; mammals; allyl compound; complementary dna; molecules; iodination; reverse transcription; chemical labeling; adenosine derivative; human; female; article; rna modifications; enzyme-assisted; mutation assays; s-adenosyl methionines; sequencing method; specific sequences
Journal Title: Journal of the American Chemical Society
Volume: 139
Issue: 48
ISSN: 0002-7863
Publisher: American Chemical Society  
Date Published: 2017-12-06
Start Page: 17213
End Page: 17216
Language: English
DOI: 10.1021/jacs.7b06837
PROVIDER: scopus
PUBMED: 29116772
PMCID: PMC5813804
Notes: Article -- Export Date: 2 January 2018 -- Source: Scopus
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MSK Authors
  1. Minkui Luo
    54 Luo