“NAD-capQ” detection and quantitation of NAD caps
- 1Department of Cell Biology and Neuroscience, Rutgers University, Piscataway, New Jersey 08854, USA
- 2Department of Genetics and Waksman Institute, Rutgers University, Piscataway, New Jersey 08854, USA
- Corresponding author: kiledjian{at}biology.rutgers.edu
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.
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Article is online at http://www.rnajournal.org/cgi/doi/10.1261/rna.067686.118.
- Received June 13, 2018.
- Accepted July 16, 2018.
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