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Interaction between composite elements in the napA promoter: both the B-box ABA-responsive complex and the RY/G complex are necessary for seed-specific expression

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Abstract

During seed maturation, the transcriptional activity of napin genes is regulated by developmental signals involving the transcriptional activator ABI3 and abscisic acid (ABA). To localize cis elements involved in the seed-specific activity of the napin napA promoter, a systematic analysis was performed focusing on two major element complexes, the B-box and RY/G. Substitution mutation analysis using promoter-reporter gene fusions in stable transgenic tobacco showed synergistic interactions between elements within these complexes. The distal part of the B-box shows similarities to abscisic acid response elements and the proximal portion contains a CA-rich element. In vitro studies involving Exonuclease III protection and electrophoretic mobility shift assays revealed binding by nuclear proteins to elements within the B-box. The distal and proximal parts of the B-box were found to bind distinct nuclear protein complexes. By gain-of-function analysis with a tetramer of the B-box fused to a truncated (−46) cauliflower mosaic virus (CaMV) 35S minimal promoter, it was demonstrated that the B-box mediates strong activity in seeds. Further, it was shown that the elements in the B-box constitute an ABA-responsive complex, since the B-box tetramer mediates ABA-responsiveness in vegetative tissues to a construct containing the CaMV virus 35S enhancer (−343 to −90). Thus, the seed-specific activity of the napA promoter relies on the combinatorial interaction between the RY/G complex and the B-box ABA-responsive complex during the ABA response in seed development.

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Authors and Affiliations

  1. Department of Plant Biology, Swedish University of Agricultural Sciences, Box 7080, 750 07, Uppsala, Sweden

    Inés Ezcurra, Mats Ellerström, Paul Wycliffe & Kjell Stålberg

  2. Department of Medical Biochemistry and Microbiology, Uppsala University, Box 575, 751 23, Uppsala, Sweden

    Lars Rask

Authors
  1. Inés Ezcurra
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  2. Mats Ellerström
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  3. Paul Wycliffe
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  4. Kjell Stålberg
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  5. Lars Rask
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Ezcurra, I., Ellerström, M., Wycliffe, P. et al. Interaction between composite elements in the napA promoter: both the B-box ABA-responsive complex and the RY/G complex are necessary for seed-specific expression. Plant Mol Biol 40, 699–709 (1999). https://doi.org/10.1023/A:1006206124512

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