"NAD-capQ" detection and quantitation of NAD caps

RNA. 2018 Oct;24(10):1418-1425. doi: 10.1261/rna.067686.118. Epub 2018 Jul 25.


RNA 5' cap structures comprising the metabolic effector nicotinamide adenine dinucleotide (NAD) have been identified in diverse organisms. Here we report a simple, two-step procedure to detect and quantitate NAD-capped RNA, termed "NAD-capQ." By use of NAD-capQ we quantitate NAD-capped RNA levels in Escherichia coli, Saccharomyces cerevisiae, and human cells, and we measure increases in NAD-capped RNA levels in cells from all three organisms harboring disruptions in their respective "deNADding" enzymes. We further show that NAD-capped RNA levels in human cells respond to changes in cellular NAD concentrations, indicating that NAD capping provides a mechanism for human cells to directly sense and respond to alterations in NAD metabolism. Our findings establish NAD-capQ as a versatile, rapid, and accessible methodology to detect and quantitate 5'-NAD caps on endogenous RNA in any organism.

Keywords: NAD cap; NAD-capQ; mRNA deNADding; mRNA decapping.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Alleles
  • Cell Line
  • Colorimetry* / methods
  • Humans
  • Intracellular Space
  • Mass Spectrometry
  • Mutation
  • NAD / chemistry*
  • NAD / metabolism
  • RNA / chemistry*
  • RNA / genetics*
  • RNA Caps / chemistry*
  • RNA Caps / genetics*
  • RNA, Messenger / chemistry
  • RNA, Messenger / genetics


  • RNA Caps
  • RNA, Messenger
  • NAD
  • RNA