Figure 5

MiSSP7 impacts the structure of the jasmonate perception complex. (a, b) MiSSP7 differentially impacts the interaction between PtJAZ6 and the two isoforms of PtMYC2 but does not alter PtJAZ6-PtJAM1.2 or PtJAZ6-PtTPR4.1 interactions. PtJAZ6 was fused to the GAL4 DBD. PtMYC2s, PtJAM1.2, PtNINJA3 and PtTPR4.1 were fused with the GAL4 AD. (c, d) MiSSP7 antagonizes dimerization between PtJAZ6 and other PtJAZ proteins. PtJAZ6 was fused to the GAL4 AD and PtJAZ proteins were fused with the GAL4 DBD. For a) and c), yeasts were transformed with MiSSP7 to give triple hybrids. TRP1, LEU2 and URA3 served as transformation markers (the latter in case of triple hybrid colonies) while HIS3 and β-galactosidase LacZ (X-Gal) were used as reporter genes. 3-Amino-1, 2, 4-triazole (3-AT) was used to suppress self-activation of the HIS3 gene. The control colonies are shown in the upper part of the panel: + is a weakly interacting pair. For b) and d), the enzymatic activity of the β-galactosidase in different yeast colonies was quantified via ONPG assay. Significant differences between yeast colonies transformed or not with MiSSP7 were assessed via pairwise Student’s t-test (*p < 0.05, **p < 0.01). White lines indicate cropped and repositioned images while the full-length blots and gels are presented in Supplementary Information online.