Arabidopsis protein <i>S</i>-acyl transferases positively mediate BR signaling through <i>S</i>-acylation of BSK1.

Abstract

Protein <i>S</i>-acyl transferases (PATs) catalyze <i>S</i>-acylation, a reversible post-translational modification critical for membrane association, trafficking, and stability of substrate proteins. Many plant proteins are potentially <i>S</i>-acylated but few have corresponding PATs identified. By using genomic editing, confocal imaging, pharmacological, genetic, and biochemical assays, we demonstrate that three Arabidopsis class C PATs positively regulate BR signaling through <i>S</i>-acylation of BRASSINOSTEROID-SIGNALING KINASE1 (BSK1). PAT19, PAT20, and PAT22 associate with the plasma membrane (PM) and the <i>trans</i>-Golgi network/early endosome (TGN/EE). Functional loss of all three genes results in a plethora of defects, indicative of reduced BR signaling and rescued by enhanced BR signaling. PAT19, PAT20, and PAT22 interact with BSK1 and are critical for the <i>S</i>-acylation of BSK1, and for BR signaling. The PM abundance of BSK1 was reduced by functional loss of <i>PAT19</i>, <i>PAT20</i>, and <i>PAT22</i> whereas abolished by its <i>S</i>-acylation-deficient point mutations, suggesting a key role of <i>S</i>-acylation in its PM targeting. Finally, an active BR analog induces vacuolar trafficking and degradation of PAT19, PAT20, or PAT22, suggesting that the <i>S</i>-acylation of BSK1 by the three PATs serves as a negative feedback module in BR signaling.