NAD⁺-capped RNAs are widespread in the Arabidopsis transcriptome and can probably be translated

As the most common RNA cap in eukaryotes, the 7-methylguanosine (m⁷G) cap impacts nearly all processes that a messenger RNA undergoes, such as splicing, polyadenylation, nuclear export, translation, and degradation. The metabolite and redox agent, nicotinamide adenine diphosphate (NAD⁺), can be used as an initiating nucleotide in RNA synthesis to result in NAD⁺-capped RNAs. Such RNAs have been identified in bacteria, yeast, and human cells, but it is not known whether they exist in plant transcriptomes. The functions of the NAD⁺ cap in RNA metabolism or translation are still poorly understood. Here, through NAD captureSeq, we show that NAD⁺-capped RNAs are widespread in Arabidopsis thaliana. NAD⁺-capped RNAs are predominantly messenger RNAs encoded by the nuclear and mitochondrial genomes, but not the chloroplast genome. NAD⁺-capped transcripts from the nuclear genome appear to be spliced and polyadenylated. Furthermore, although NAD⁺-capped transcripts constitute a small proportion of the total transcript pool from any gene, they are enriched in the polysomal fraction and associate with translating ribosomes. Our findings implicate the existence of as yet unknown mechanisms whereby the RNA NAD⁺ cap interfaces with RNA metabolic processes as well as translation initiation. More importantly, our findings suggest that cellular metabolic and/or redox states may influence, or be regulated by, mRNA NAD⁺ capping.