Co-Authors:
Meller, Y., Department of Plant Genetics, Weizmann Institute of Science, Rehovot, 76100, Israel
Sessa, G., Department of Plant Genetics, Weizmann Institute of Science, Rehovot, 76100, Israel
Eyal, Y., Department of Plant Genetics, Weizmann Institute of Science, Rehovot, 76100, Israel
Fluhr, R., Department of Plant Genetics, Weizmann Institute of Science, Rehovot, 76100, Israel
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. © 1993 Kluwer Academic Publishers.
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