PLX251563

GSE90164: Nm-seq finds thousands of modified 2-O-methylation sites in mRNA with base precision

• Organsim human
• Type RNASEQ
• Target gene
• Project ARCHS4

Chemical modifications of messenger RNA (mRNA) molecules, collectively known as the epitranscriptome, regulate gene expression and currently comprise only base modifications. Whereas bases can be modified at every position, the ribose can solely be modified at the 2 position with methyl as the only known natural group. 2-O-methylation is an abundant modification in rRNA, tRNA and in the 5-cap of mRNA, important for multiple aspects of translation fidelity and innate immunity. Despite recent advances in high-throughput detection, its inert chemical nature still limits sensitivity and precludes detection of Nm present in rare RNA molecules or at low stoichiometry. We leveraged the differential sensitivity of 2-O-methylated and 2-hydroxylated nucleosides to periodate oxidation to develop Nm-seq, a highly sensitive method for mapping of 2-O-methylation in the transcriptome with base precision. Nm-seq exposed thousands of sites in human mRNA, the majority of which occurred in just a few codons, in line with a characteristic motif, and clustered around splice junctions. Our discovery adds yet another modification to the known repertoire and lays the foundation for future functional investigations as to the role of Nm in translation, splicing, regulated protein binding and more. SOURCE: Sharon Moshitch-Moshkovitz (sharon.moshkovitz@sheba.health.gov.il) - Sheba Medical Center