Accurate detection of m6A RNA modifications in native RNA sequences

Nat Commun. 2019 Sep 9;10(1):4079. doi: 10.1038/s41467-019-11713-9.

Abstract

The epitranscriptomics field has undergone an enormous expansion in the last few years; however, a major limitation is the lack of generic methods to map RNA modifications transcriptome-wide. Here, we show that using direct RNA sequencing, N6-methyladenosine (m6A) RNA modifications can be detected with high accuracy, in the form of systematic errors and decreased base-calling qualities. Specifically, we find that our algorithm, trained with m6A-modified and unmodified synthetic sequences, can predict m6A RNA modifications with ~90% accuracy. We then extend our findings to yeast data sets, finding that our method can identify m6A RNA modifications in vivo with an accuracy of 87%. Moreover, we further validate our method by showing that these 'errors' are typically not observed in yeast ime4-knockout strains, which lack m6A modifications. Our results open avenues to investigate the biological roles of RNA modifications in their native RNA context.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Adenosine / analogs & derivatives*
  • Adenosine / metabolism
  • Base Sequence
  • Electricity
  • RNA / genetics*
  • RNA / metabolism*
  • Saccharomyces cerevisiae / genetics
  • Sequence Analysis, RNA
  • Support Vector Machine

Substances

  • RNA
  • N-methyladenosine
  • Adenosine