Combinatorial interactions of multiple cis-elements regulating the induction of the Arabidopsis XERO2 dehydrin gene by abscisic acid and cold.

Multiple combinations of mutations in the promoter of the XERO2 dehydrin gene were used to identify elements involved in ABA and cold induction. Mutating one of the three ACGT elements (ACGT1) increases expression in the absence of cold or ABA. An AT-rich element is a novel partner (coupling element) of ACGT-containing ABA-responsive cis-elements. A 12-bp palindrome also acts as a coupling element for ABA induction and includes one of the three dehydration-responsive element/C-repeat (DRE/CRT) elements and two overlapping motifs (TGTCG and TCGGC) previously shown to be statistically enriched in ABA-dependent and 'VP1 or ABA'-dependent activated genes (Plant Physiol. 2005; 139:437). At least two of the DRE/CRT elements are required for significant cold induction. During cold induction the AT-rich element also functions as a coupling element and ACGT1 is involved in repressing this induction. Two of the ACGT and DRE/CRT elements overlap, and mutating a single base in the ACGT of either of the two GCCGACGT sequences while retaining a DRE element reduced both ABA and cold induction. Changing the spatial relationships between the elements by deletion, inversion or insertion of DNA sequences reduced both cold and ABA induction. Overexpression of CBF1, -2 or -3 induced XERO2 expression in untreated plants. The ABI5 transcription factor may have a role in ABA-induced XERO2 expression, whereas ABI3 and ABI4 do not. The GCA2 gene was essential for both cold and ABA induction. A combination of the same overlapping and shared elements is used in the regulation of transcription by ABA and cold.