MePMe-seq: antibody-free simultaneous m⁶A and m⁵C mapping in mRNA by metabolic propargyl labeling and sequencing.

ABSTRACT: Internal modifications of mRNA have emerged as widespread and versatile regulatory mechanism to control gene expression at the post-transcriptional level. Most of these modifications are methyl groups, making S-adenosyl-L-methionine (SAM) a central metabolic hub. Here we show that metabolic labeling with a clickable metabolic precursor of SAM, propargyl-selenohomocysteine (PSH), enables detection and identification of various methylation sites. Propargylated A, C, and G nucleosides form at detectable amounts via intracellular generation of the corresponding SAM analogue. Integration into next generation sequencing enables mapping of N⁶-methyladenosine (m⁶A) and 5-methylcytidine (m⁵C) sites in mRNA with single nucleotide precision (MePMe-seq). Analysis of th

SUBMITTER: Hartstock K 

PROVIDER: S-EPMC10630376 | biostudies-literature | 2023 Nov

REPOSITORIES: biostudies-literature