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Published on May 11, 2007; 10.1104/pp.107.096115

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Received January 18, 2007
Accepted May 3, 2007

Identification of a Bipartite Jasmonate-responsive Promoter Element in the Periwinkle ORCA3 Transcription Factor Gene which Interacts Specifically with AT-hook DNA-binding Proteins

Débora Vom Endt , Marina Soares e Silva , Jan W. Kijne , Giancarlo Pasquali , and Johan Memelink *

Institute of Biology, Leiden University, Clusius Laboratory, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands; Centro de Biotecnologia/UFRGS, Av. Bento Gonçalves, 9500 Prédio 43431, C.P. 15005, CEP 91501-970, Porto Alegre, RS, Brazil

* Corresponding author; email: j.memelink{at}biology.leidenuniv.nl.

Jasmonates are plant signalling molecules that play key roles in defense against certain pathogens and insects, among others by controlling the biosynthesis of protective secondary metabolites. In Catharanthus roseus, the AP2-domain transcription factor ORCA3 is involved in the jasmonate-responsive activation of terpenoid indole alkaloid biosynthetic genes. ORCA3 gene expression is itself induced by jasmonate. By loss- and gain-of-function experiments we located a 74 bp region within the ORCA3 promoter, which contains an autonomous jasmonate-responsive element (JRE). The ORCA3 JRE is composed of two important sequences, a quantitative sequence responsible for a high level of expression, and a qualitative sequence that appears to act as an on/off switch in response to methyl-jasmonate (MeJA). We isolated 12 different DNA-binding proteins having one of four different types of DNA-binding domains, using the ORCA3 JRE as bait in a yeast one-hybrid transcription factor screening. The binding of one class of proteins bearing a single AT-hook DNA-binding motif was affected by mutations in the quantitative sequence within the JRE. Two of the AT-hook proteins tested had a weak activating effect on JRE-mediated reporter gene expression, suggesting that AT-hook family members may be involved in determining the level of expression of ORCA3 in response to jasmonate.






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