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Two novel AP2/ERF domain proteins interact with cis ‐element VWRE for wound‐induced expression of the Tobacco tpoxN1 gene
Author(s) -
Sasaki Katsutomo,
Mitsuhara Ichiro,
Seo Shigemi,
Ito Hiroyuki,
Matsui Hirokazu,
Ohashi Yuko
Publication year - 2007
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/j.1365-313x.2007.03111.x
Subject(s) - transcription factor , gene , subfamily , biology , gene expression , microbiology and biotechnology , homology (biology) , genetics , two hybrid screening
Summary The vascular system‐specific and wound‐responsive cis ‐element (VWRE) has been identified as a novel cis ‐element for wound‐induced and vascular system‐specific expression of the tobacco peroxidase gene, tpoxN1 . Here we isolated two independent clones that encode VWRE binding proteins by yeast one‐hybrid screening. As the gene products have an AP2/ERF (APETALA2/ethylene‐responsive factor) domain, and the transcripts were accumulated transiently after wounding, we named them wound‐responsive AP2/ERF‐like factor 1 (WRAF1) and WRAF2. The AP2/ERF domains of the two WRAFs share 97% homology, and are classified into the ERF subfamily B‐4. Gel mobility shift analysis indicated that WRAFs specifically bind VWRE, which contains no known cis ‐elements for other AP2/ERF proteins. The binding activity of the WRAFs was found to be localized in the AP2/ERF domain. The WRAFs transactivated a promoter containing four tandem repeats of the VWRE, but not that of the mutated VWRE. Overexpression of the WRAF genes led to constitutive expression of the potential target gene, tpoxN1 , in unwounded transgenic plants. These results indicate that the novel transcription factors WRAF1 and WRAF2 bind the VWRE as positive regulators for the expression of the tpoxN1 gene.

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