Nonsense-mediated decay factors are involved in the regulation of selenoprotein mRNA levels during selenium deficiency

Abstract

Selenoproteins contain the unique amino acid selenocysteine (Sec), which is encoded by the triplet UGA. Since UGA also serves as a stop codon, it has been postulated that selenoprotein mRNAs are targeted for degradation by the nonsense-mediated mRNA decay pathway (NMD). Several reports have observed a hierarchy of selenoprotein mRNA expression when selenium (Se) is limiting, whereby the abundance of certain transcripts decline while others do not. We sought to investigate the role of NMD in this hierarchical response that selenoprotein mRNAs exhibit to environmental Se status. Selenoprotein mRNAs were categorized as being predicted sensitive or resistant to NMD based on the requirements held by the current model. About half of the selenoprotein transcriptome was predicted to be sensitive to NMD and showed significant changes in mRNA abundance in response to cellular Se status. The other half that was predicted to be resistant to NMD did not respond to Se status. RNA immunoprecipitation with essential NMD factor UPF1 revealed that the mRNAs that were the most sensitive to Se status were also the most enriched on UPF1 during Se deficiency. Furthermore, depletion of SMG1, the kinase responsible for UPF1 phosphorylation and NMD activation, abrogated the decline in transcript abundance of Se-responsive transcripts. Lastly, mRNA decay rates of Se-responsive transcripts were altered upon the addition of Se to resemble the slower decay rates of nonresponsive transcripts. Taken together, these results present novel evidence in support of a crucial role for the NMD pathway in regulating selenoprotein mRNA levels when Se is limiting.