The RALF1 peptide-FERONIA complex phosphorylates the endosomal sorting protein FREE1 to attenuate abscisic acid signaling

Abstract The receptor-like kinase FERONIA (FER), together with its ligand rapid alkalinization factor 1 (RALF1) peptide, plays a crucial role in regulating stress responses, including its involvement in modulating abscisic acid (ABA) signaling. FER has been shown to activate ABA insensitive 2 in the cytoplasm, leading to the suppression of ABA signaling. However, its regulation of nucleus events in the ABA response remains unclear. FREE1, identified as a plant-specific component of the endosomal sorting complex required for transport in eukaryotes, serves as an important negative regulator in ABA signaling. In this study, we elucidate that upon RALF1 treatment, FER phosphorylates FREE1, promoting the accumulation of FREE1 protein in the nucleus in Arabidopsis (Arabidopsis thaliana). Consequently, FREE1 suppresses ABA sensitivity by inhibiting the expression of ABA-response genes. Mutating the 6 identified phosphorylation sites on FREE1, mediated by FER, to nonphosphorylable residues results in reduced nucleus localization of FREE1 and increased hypersensitivity to ABA. Our data also show that these 6 phosphorylation sites are likely involved in regulating plant survival under salt stress. Collectively, our study not only unveils an additional function of FER in attenuating ABA signaling in the nucleus but also provides a possible insight into the role of the RALF1-FER-FREE1 module in coordinating plant growth and salt stress tolerance.