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“NAD-capQ” detection and quantitation of NAD caps

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...

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Autores principales: Grudzien-Nogalska, Ewa, Bird, Jeremy G., Nickels, Bryce E., Kiledjian, Megerditch
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6140466/
https://www.ncbi.nlm.nih.gov/pubmed/30045887
http://dx.doi.org/10.1261/rna.067686.118
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author Grudzien-Nogalska, Ewa
Bird, Jeremy G.
Nickels, Bryce E.
Kiledjian, Megerditch
author_facet Grudzien-Nogalska, Ewa
Bird, Jeremy G.
Nickels, Bryce E.
Kiledjian, Megerditch
author_sort Grudzien-Nogalska, Ewa
collection PubMed
description 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.
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spelling pubmed-61404662019-10-01 “NAD-capQ” detection and quantitation of NAD caps Grudzien-Nogalska, Ewa Bird, Jeremy G. Nickels, Bryce E. Kiledjian, Megerditch RNA Method 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. Cold Spring Harbor Laboratory Press 2018-10 /pmc/articles/PMC6140466/ /pubmed/30045887 http://dx.doi.org/10.1261/rna.067686.118 Text en © 2018 Grudzien-Nogalska et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
spellingShingle Method
Grudzien-Nogalska, Ewa
Bird, Jeremy G.
Nickels, Bryce E.
Kiledjian, Megerditch
“NAD-capQ” detection and quantitation of NAD caps
title “NAD-capQ” detection and quantitation of NAD caps
title_full “NAD-capQ” detection and quantitation of NAD caps
title_fullStr “NAD-capQ” detection and quantitation of NAD caps
title_full_unstemmed “NAD-capQ” detection and quantitation of NAD caps
title_short “NAD-capQ” detection and quantitation of NAD caps
title_sort “nad-capq” detection and quantitation of nad caps
topic Method
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6140466/
https://www.ncbi.nlm.nih.gov/pubmed/30045887
http://dx.doi.org/10.1261/rna.067686.118
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