Transcriptional responses to nitric oxide are widescale and source-dependent.

The transcriptomic effects of nitric oxide (NO) have been widely studied across phylogeny. However, while gene expression is canonically altered by NO, general principles have not emerged. Here, we characterize genetic regulation within a single cell type after exposure to NO derived from endogenous or synthetic donor compounds or produced by 3 different NO synthase (NOS) isoforms under basal and activated conditions. Using RNAseq, we uncover distinct, source-dependent effects of NO on as many as ∼10,000 genes mediated largely by S-nitrosylation. NOS enzymes and NO donors each generated unique transcriptional responses. Our data reveal non-overlapping transcriptional responses to NO that are likely mediated by distinct effectors and enzymes and highlight that NO-treated cell systems may undergo a dramatic and widespread transcriptional response.