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J. Biol. Chem., Vol. 279, Issue 53, 55770-55779, December 31, 2004

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A WRKY Gene from Creosote Bush Encodes an Activator of the Abscisic Acid Signaling Pathway^*

Xiaolu Zou, Jeffrey R. Seemann, Dawn Neuman, and Qingxi J. Shen¶

From the Department of Biological Sciences, University of Nevada, Las Vegas, Nevada 89154 and the Department of Cell and Molecular Biology, University of Rhode Island, Kingston, Rhode Island 02881

The creosote bush (Larrea tridentata) is a xerophytic evergreen C3 shrub thriving in vast arid areas of North America. As the first step toward understanding the molecular mechanisms controlling the drought tolerance of this desert plant, we have isolated a dozen genes encoding transcription factors, including LtWRKY21 that encodes a protein of 314 amino acid residues. Transient expression studies with the GFP-LtWRKY21 fusion construct indicate that the LtWRKY21 protein is localized in the nucleus and is able to activate the promoter of an abscisic acid (ABA)-inducible gene, HVA22, in a dosage-dependent manner. The transactivating activity of LtWRKY21 relies on the C-terminal sequence containing the WRKY domain and a N-terminal motif that is essential for the repression activity of some regulators in ethylene signaling. LtWRKY21 interacts synergistically with ABA and transcriptional activators VP1 and ABI5 to control the expression of the HVA22 promoter. Co-expression of VP1, ABI5, and LtWRKY21 leads to a much higher expression of the HVA22 promoter than does the ABA treatment alone. In contrast, the Lt-WRKY21-mediated transactivation is inhibited by two known negative regulators of ABA signaling: 1-butanol, an inhibitor of phospholipase D, and abi1-1, a dominant negative mutant protein phosphatase. Interestingly, abi1-1 does not block the synergistic effect of LtWRKY21, VP1, and ABI5 co-expression, indicating that LtWRKY21, VP1, and ABI5 may form a complex that functions downstream of ABI1 to control ABA-regulated expression of genes.

Received for publication, July 28, 2004

The nucleotide sequence(s) reported in this paper has been submitted to the DDBJ/GenBank^TM/EBI Data Bank with accession number(s) AY792618.

^* This work was supported by a University of Nevada, Las Vegas start-up grant, National Institutes of Health Biomedical Research Infrastructure Network Seed Grant P20 RR16464, and National Science Foundation Experimental Program to Stimulate Competitive Research (EPSCoR) Seed Grant EPS-9977809 (to Q. J. S.) and National Science Foundation EPSCoR IAAS Fellowship EPS-0132556 (to X. Z.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ To whom correspondence should be addressed: Dept. of Biological Sciences, M/S 4004, 4505 Maryland Parkway, Las Vegas, NV 89154. Tel.: 702-895-4704; Fax: 702-895-3956; E-mail: jeffery.shen{at}ccmail.nevada.edu.

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