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  • Messenger RNA 5' NAD+ Capping Is a Dynamic Regulatory Epitranscriptome Mark That Is Required for Proper Response to Abscisic Acid in Arabidopsis.

Messenger RNA 5' NAD+ Capping Is a Dynamic Regulatory Epitranscriptome Mark That Is Required for Proper Response to Abscisic Acid in Arabidopsis.

Developmental cell (2020-12-09)
Xiang Yu, Matthew R Willmann, Lee E Vandivier, Sophie Trefely, Marianne C Kramer, Jeffrey Shapiro, Rong Guo, Eric Lyons, Nathaniel W Snyder, Brian D Gregory
ABSTRACT

Although eukaryotic messenger RNAs (mRNAs) normally possess a 5' end N7-methyl guanosine (m7G) cap, a non-canonical 5' nicotinamide adenine dinucleotide (NAD+) cap can tag certain transcripts for degradation mediated by the NAD+ decapping enzyme DXO1. Despite this importance, whether NAD+ capping dynamically responds to specific stimuli to regulate eukaryotic transcriptomes remains unknown. Here, we reveal a link between NAD+ capping and tissue- and hormone response-specific mRNA stability. In the absence of DXO1 function, transcripts displaying a high proportion of NAD+ capping are instead processed into RNA-dependent RNA polymerase 6-dependent small RNAs, resulting in their continued turnover likely to free the NAD+ molecules. Additionally, the NAD+-capped transcriptome is significantly remodeled in response to the essential plant hormone abscisic acid in a mechanism that is primarily independent of DXO1. Overall, our findings reveal a previously uncharacterized and essential role of NAD+ capping in dynamically regulating transcript stability during specific physiological responses.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
2-Propanol, suitable for HPLC, 99.9%
Sigma-Aldrich
Phenol – chloroform – isoamyl alcohol mixture, BioUltra, for molecular biology, 25:24:1
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(+)-Sodium L-ascorbate, crystalline, ≥98%
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β-Nicotinamide adenine dinucleotide hydrate, ≥96.5% (HPLC), ≥96.5% (spectrophotometric assay), from yeast
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Chloroform:Isoamyl alcohol 24:1, suitable for nucleic acid purification
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Copper(II) sulfate, ReagentPlus®, ≥99%
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HEPES sodium salt, ≥99.5% (titration)
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Tris(3-hydroxypropyltriazolylmethyl)amine, 95%
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Sodium acetate solution, BioUltra, for molecular biology, ~3 M in H2O
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4-Pentyn-1-ol, 97%
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(+)-Abscisic acid, ≥98% (HPLC)
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Albumin, Acetylated from bovine serum, protease free, for molecular biology
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