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DNA-protein interactions on a cis-DNA element essential for ethylene regulation

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Abstract

The PRB-1b gene encodes for a basic-type component of the pathogenesis-related PR-1 protein family. In leaves of tobacco plants, PRB-1b mRNA accumulation is rapidly induced by the application of exogenous ethylene. Promoter deletion analysis was performed in transgenic tobacco plants to delineate cis-acting elements necessary for ethylene responsiveness of the PRB-1b gene. The promoter sequence from position −213 was sufficient to enhance a 20 fold increase of β-glucuronidase reporter gene expression in transgenic tobacco leaves exposed to 20 μl/l of ethylene, however −141 bp were not. The functional study was correlated with in vitro analysis of the nuclear protein-DNA complexes formed on the promoter element identified as necessary for ethylene induction. Gel-shift analysis using restriction fragments spanning the sequence between position −237 and −143 revealed two distinct nuclear protein-DNA interactions. The protein-binding sequences were mapped to the contiguous regions G (−200 to −178) and Y (−179 to −154) by gel-shift analysis using oligonucleotides. Fractionation of crude nuclear extract by heparin-agarose chromatography resulted in the differential elution of the two binding activities. The DNA-nuclear protein interactions characterized in vitro can be part of the molecular events which mediate the transcriptional regulation of the PRB-1b gene by ethylene.

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Authors and Affiliations

  1. Department of Plant Genetics, Weizmann Institute of Science, 76100, Rehovot, Israel

    Yael Meller, Guido Sessa, Yoram Eyal & Robert Fluhr

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  1. Yael Meller
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  2. Guido Sessa
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  3. Yoram Eyal
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  4. Robert Fluhr
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Meller, Y., Sessa, G., Eyal, Y. et al. DNA-protein interactions on a cis-DNA element essential for ethylene regulation. Plant Mol Biol 23, 453–463 (1993). https://doi.org/10.1007/BF00019294

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  • DOI: https://doi.org/10.1007/BF00019294

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