Capturing the interactome of newly transcribed RNA

Nat Methods. 2018 Mar;15(3):213-220. doi: 10.1038/nmeth.4595. Epub 2018 Feb 12.

Abstract

We combine the labeling of newly transcribed RNAs with 5-ethynyluridine with the characterization of bound proteins. This approach, named capture of the newly transcribed RNA interactome using click chemistry (RICK), systematically captures proteins bound to a wide range of RNAs, including nascent RNAs and traditionally neglected nonpolyadenylated RNAs. RICK has identified mitotic regulators amongst other novel RNA-binding proteins with preferential affinity for nonpolyadenylated RNAs, revealed a link between metabolic enzymes/factors and nascent RNAs, and expanded the known RNA-bound proteome of mouse embryonic stem cells. RICK will facilitate an in-depth interrogation of the total RNA-bound proteome in different cells and systems.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Click Chemistry / methods*
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / metabolism
  • HeLa Cells
  • High-Throughput Nucleotide Sequencing / methods
  • Humans
  • Mass Spectrometry / methods
  • Mice
  • Protein Interaction Maps
  • Proteome / metabolism*
  • RNA / genetics
  • RNA / metabolism*
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism*
  • Uridine / analogs & derivatives
  • Uridine / chemistry

Substances

  • Proteome
  • RNA-Binding Proteins
  • RNA
  • Uridine