Ribosome stalling-induced <i>NIP5;1</i> mRNA decay triggers ARGONAUTE1-dependent transcription downregulation

In eukaryotes, messenger RNA (mRNA) accumulation is regulated through the levels of transcription, processing, and degradation. Here, we uncover the multi-level regulatory mechanism governing the expression of NIP5;1, a boron (B) diffusion facilitator in Arabidopsis. B-dependent NIP5;1 mRNA degradation is triggered by ribosome stalling at an AUGUAA sequence in its 5'-untranslated region. We showed that deletion of ATGTAA also abolishes B-dependent transcriptional downregulation, revealing a dual role of this sequence in both mRNA degradation and transcriptional control. Small RNAs (sRNAs) and ARGONAUTE1 (AGO1) are implicated in mRNA-degradation-mediated B-dependent transcriptional downregulation: a 5'-3' exonuclease mutant, xrn4, presents both elevated levels of NIP5;1 mRNA degradation intermediates and transcriptional downregulation; AGO1-associated sRNA-sequencing reveals the presence of sRNAs with sequences upstream of NIP5;1 AUGUAA; and nascent mRNA profiling by global run-on sequencing demonstrates RNA polymerase II pausing at ATGTAA, a phenomenon diminished in the ago1 mutant that lacks B-dependent transcriptional downregulation. These findings point to multi-level coordination of NIP5;1 expression with the AUGUAA sequence at its core: ribosome stalling orchestrates translational inhibition, mRNA degradation and transcriptional downregulation in response to B. The fast response resulting from this synergy suggests that similar mechanisms may exist in other eukaryotic systems for efficient and rapid regulation of gene expression.