High-resolution N(6) -methyladenosine (m(6) A) map using photo-crosslinking-assisted m(6) A sequencing

Angew Chem Int Ed Engl. 2015 Jan 26;54(5):1587-90. doi: 10.1002/anie.201410647. Epub 2014 Dec 9.

Abstract

N(6) -methyladenosine (m(6) A) is an abundant internal modification in eukaryotic mRNA and plays regulatory roles in mRNA metabolism. However, methods to precisely locate the m(6) A modification remain limited. We present here a photo-crosslinking-assisted m(6) A sequencing strategy (PA-m(6) A-seq) to more accurately define sites with m(6) A modification. Using this strategy, we obtained a high-resolution map of m(6) A in a human transcriptome. The map resembles the general distribution pattern observed previously, and reveals new m(6) A sites at base resolution. Our results provide insight into the relationship between the methylation regions and the binding sites of RNA-binding proteins.

Keywords: N6-methyladenosine; RNA modification; photo-crosslinking; transcriptome sequencing.

Publication types

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

MeSH terms

  • Adenine / analogs & derivatives*
  • Adenine / analysis
  • Adenine / immunology
  • Antibodies / immunology
  • HeLa Cells
  • Humans
  • Polymerase Chain Reaction
  • RNA / chemistry*
  • RNA / metabolism
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism
  • Sequence Analysis, RNA
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Thiourea / chemistry
  • Thiourea / metabolism
  • Ultraviolet Rays

Substances

  • Antibodies
  • RNA-Binding Proteins
  • N(6)-methoxyadenine
  • RNA
  • Thiourea
  • Adenine