Abstract
A highly active synthetic auxin response element (AuxRE), referred to as DR5, was created by performing site-directed mutations in a natural composite AuxRE found in the soybean GH3 promoter. DR5 consisted of tandem direct repeats of 11 bp that included the auxin-responsive TGTCTC element. The DR5 AuxRE showed greater auxin responsiveness than a natural composite AuxRE and the GH3 promoter when assayed by transient expression in carrot protoplasts or in stably transformed Arabidopsis seedlings, and it provides a useful reporter gene for studying auxin-responsive transcription in wild-type plants and mutants. An auxin response transcription factor, ARF1, bound with specificity to the DR5 AuxRE in vitro and interacted with Aux/IAA proteins in a yeast two-hybrid system. Cotransfection experiments with natural and synthetic AuxRE reporter genes and effector genes encoding Aux/IAA proteins showed that overexpression of Aux/IAA proteins in carrot protoplasts resulted in specific repression of TGTCTC AuxRE reporter gene expression.
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- Abel S., Nguyen M. D., Theologis A. The PS-IAA4/5-like family of early auxin-inducible mRNAs in Arabidopsis thaliana. J Mol Biol. 1995 Aug 25;251(4):533–549. doi: 10.1006/jmbi.1995.0454. [DOI] [PubMed] [Google Scholar]
- Abel S., Oeller P. W., Theologis A. Early auxin-induced genes encode short-lived nuclear proteins. Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):326–330. doi: 10.1073/pnas.91.1.326. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Abel S., Theologis A. Early genes and auxin action. Plant Physiol. 1996 May;111(1):9–17. doi: 10.1104/pp.111.1.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ainley W. M., Walker J. C., Nagao R. T., Key J. L. Sequence and characterization of two auxin-regulated genes from soybean. J Biol Chem. 1988 Aug 5;263(22):10658–10666. [PubMed] [Google Scholar]
- Bevan M. Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res. 1984 Nov 26;12(22):8711–8721. doi: 10.1093/nar/12.22.8711. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Durfee T., Becherer K., Chen P. L., Yeh S. H., Yang Y., Kilburn A. E., Lee W. H., Elledge S. J. The retinoblastoma protein associates with the protein phosphatase type 1 catalytic subunit. Genes Dev. 1993 Apr;7(4):555–569. doi: 10.1101/gad.7.4.555. [DOI] [PubMed] [Google Scholar]
- Hagen G., Martin G., Li Y., Guilfoyle T. J. Auxin-induced expression of the soybean GH3 promoter in transgenic tobacco plants. Plant Mol Biol. 1991 Sep;17(3):567–579. doi: 10.1007/BF00040658. [DOI] [PubMed] [Google Scholar]
- Leyser H. M., Lincoln C. A., Timpte C., Lammer D., Turner J., Estelle M. Arabidopsis auxin-resistance gene AXR1 encodes a protein related to ubiquitin-activating enzyme E1. Nature. 1993 Jul 8;364(6433):161–164. doi: 10.1038/364161a0. [DOI] [PubMed] [Google Scholar]
- Liu Z. B., Ulmasov T., Shi X., Hagen G., Guilfoyle T. J. Soybean GH3 promoter contains multiple auxin-inducible elements. Plant Cell. 1994 May;6(5):645–657. doi: 10.1105/tpc.6.5.645. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McClure B. A., Hagen G., Brown C. S., Gee M. A., Guilfoyle T. J. Transcription, organization, and sequence of an auxin-regulated gene cluster in soybean. Plant Cell. 1989 Feb;1(2):229–239. doi: 10.1105/tpc.1.2.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Oeller P. W., Keller J. A., Parks J. E., Silbert J. E., Theologis A. Structural characterization of the early indoleacetic acid-inducible genes, PS-IAA4/5 and PS-IAA6, of pea (Pisum sativum L.). J Mol Biol. 1993 Oct 20;233(4):789–798. doi: 10.1006/jmbi.1993.1555. [DOI] [PubMed] [Google Scholar]