JAZs are acetylation substrates of HopZ1a.

(A) HopZ1a acetylates GmJAZ1 in vitro. Tag-free HopZ1a and HopZ1a(C216A), and HIS-GmJAZ1 were purified from E. coli and subjected to in vitro acetylation assays. The acetylated proteins were detected by autoradiography after exposure at −80°C for five days. (B) HopZ1a acetylates MBP-AtJAZ6-HIS in vitro. (C) The mutant AtJAZ6ΔJas no longer interacts with HopZ1a. Purified HopZ1a was incubated with MBP-AtJAZ6-HIS or MBP-AtJAZ6ΔJas-HIS in the in vitro pull-down assay. (D) HopZ1a no longer acetylates AtJAZ6ΔJas. (E) HopZ1a does not trigger the degradation of AtJAZ6ΔJas. AtJAZ6ΔJas-YFP-HA and 3×FLAG-HopZ1a were co-expressed in N. benthamiana. The abundance of AtJAZ6ΔJas was detected by western blots. All the in vitro acetylation experiments were repeated at least three times with similar results. The in vitro pull-down and degradation experiments were repeated twice with similar results.