"NAD-capQ" detection and quantitation of NAD caps

Ewa Grudzien-Nogalska, Jeremy G. Bird, Bryce Nickels, Megerditch Kiledjian

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1418-1425
Number of pages8
JournalRNA
Volume24
Issue number10
DOIs
Publication statusPublished - Oct 2018

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All Science Journal Classification (ASJC) codes

  • Molecular Biology

Keywords

  • MRNA deNADding
  • MRNA decapping
  • NAD cap
  • NAD-capQ

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