The jasmonate-inducible AP2/ERF-domain transcription factor ORCA3 activates gene expression via interaction with a jasmonate-responsive promoter element

The AP2/ERF-domain transcription factor ORCA3 is a master regulator of primary and secondary metabolism in Catharanthus roseus (periwinkle). Here we demonstrate that ORCA3 specifically binds to and activates gene expression via a previously characterized jasmonate- and elicitor-responsive element (JERE) in the promoter of the terpenoid indole alkaloid biosynthetic gene Strictosidine synthase (Str). Functional characterization of different domains in the ORCA3 protein in yeast and plant cells revealed the presence of an N-terminal acidic activation domain and a serine-rich C-terminal domain with a negative regulatory function. Orca3 mRNA accumulation was rapidly induced by the plant stress hormone methyljasmonate with biphasic kinetics. A precursor and an intermediate of the jasmonate biosynthetic pathway also induced Orca3 gene expression, further substantiating the role for ORCA3 in jasmonate signaling. The protein synthesis inhibitor cycloheximide did not inhibit jasmonate-responsive expression of Orca3, nor of its target genes Str and Tryptophan decarboxylase (Tdc). In conclusion, ORCA3 regulates jasmonate-responsive expression of the Str gene via direct interaction with the JERE. The activating activities of ORCA proteins do not seem to depend on jasmonate-induced de novo protein synthesis, but presumably occur via modification of pre-existing ORCA protein.