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Cloning of a DNA-binding protein that interacts with the ethylene-responsive enhancer element of the carnation GST1 gene

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Abstract

Ethylene transcriptionally activates a glutathione S-transferase gene (GST1) at the onset of the senescence program in carnation (Dianthus caryophyllus L.) flower petals. A 126 bp region of the GST1 promoter sequence has been identified as an ethylene-responsive enhancer element (ERE). In this paper, we demonstrate the ability of nuclear proteins from senescing petals to recognize a 22 bp sequence within the ERE (ERE oligonucleotide). Mutation of the ERE oligonucleotide sequence significantly alters the strength of this nuclear protein-DNA association. The wild-type ERE oligonucleotide sequence was used to isolate a cDNA clone encoding a sequence-specific DNA binding protein. Nucleotide sequencing and deduced amino acid sequence analysis of this cDNA predicted a 32 kDa protein which we have designated carnation ethylene-responsive element-binding protein-1 (CEBP-1). The mRNA expression pattern of CEBP-1 suggests that it is not transcriptionally regulated by ethylene. The amino acid sequence homology of CEBP-1 with other plant nucleic acid binding proteins indicates a conserved nucleic acid binding domain. Within this domain are two highly conserved RNA-binding motifs, RNP-1 and RNP-2. An acidic region and a putative nuclear localization signal are also identified.

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Abbreviations

ERE:

ethylene-responsive element

GST:

glutathione S-transferase

RNP:

ribonucleoprotein

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Authors and Affiliations

  1. Department of Horticulture, Purdue University, 47907-1165, West Lafayette, IN, USA

    Julie M. Maxson & William R. Woodson

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  1. Julie M. Maxson
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  2. William R. Woodson
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Maxson, J.M., Woodson, W.R. Cloning of a DNA-binding protein that interacts with the ethylene-responsive enhancer element of the carnation GST1 gene. Plant Mol Biol 31, 751–759 (1996). https://doi.org/10.1007/BF00019463

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  • DOI: https://doi.org/10.1007/BF00019463

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