The m⁶ A reader ECT2 post-transcriptionally regulates proteasome activity in Arabidopsis

Methylation of internal adenosine at nitrogen-6 position (m⁶ A) is the most abundant post-transcriptional modification in eukaryotic RNAs. These modifications are recognized by m⁶ A-binding proteins ('readers') that affect downstream functions. In plants, the scope of gene expression regulation by reader proteins is not clear. Here, overexpression and loss-of-function mutants were used to characterize the role of the Arabidopsis m⁶ A reader ECT2 in proteasome regulation. ECT2 regulates the mRNA levels of the proteasome regulator PTRE1 and of several 20S proteasome subunits, resulting in enhanced 26S proteasome activity. This regulation is dependent on ECT2 m⁶ A binding function. Interestingly, though ECT2 positively regulates proteasome activity in both young and mature plants, PTRE1 has different regulatory effects in different developmental stages. In mature plants, PTRE1 inhibits 26S proteasome activity, while in seedlings PTRE1 knockout mutants have reduced 26S proteasome activity. Taken together, our results suggest a novel epitranscriptomic mechanism of proteasome regulation by ECT2 that is used to fine tune proteasome activity by affecting the expression of PTRE1 and 20S proteasome subunits.