Hydrogen sulfide improves salt tolerance through persulfidation of PMA1 in Arabidopsis

A new interaction was found between PMA1 and GRF4. H₂S promotes the interaction through persulfidated Cys446 of PMA1. H₂S activates PMA1 to maintain K⁺/Na⁺ homeostasis through persulfidation under salt stress. Plasma membrane H⁺-ATPase (PMA) is a transmembrane transporter responsible for pumping protons, and its contribution to salt resistance is indispensable in plants. Hydrogen sulfide (H₂S), a small signaling gas molecule, plays the important roles in facilitating adaptation of plants to salt stress. However, how H₂S regulates PMA activity remains largely unclear. Here, we show a possible original mechanism for H₂S to regulate PMA activity. PMA1, a predominant member in the PMA family of Arabidopsis, has a non-conservative persulfidated cysteine (Cys) residue (Cys446), which is exposed on the surface of PMA1 and located in cation transporter/ATPase domain. A new interaction of PMA1 and GENERAL REGULATORY FACTOR 4 (GRF4, belongs to the 14-3-3 protein family) was found by chemical crosslinking coupled with mass spectrometry (CXMS) in vivo. H₂S-mediated persulfidation promoted the binding of PMA1 to GRF4. Further studies showed that H₂S enhanced instantaneous H⁺ efflux and maintained K⁺/Na⁺ homeostasis under salt stress. In light of these findings, we suggest that H₂S promotes the binding of PMA1 to GRF4 through persulfidation, and then activating PMA, thus improving the salt tolerance of Arabidopsis.