Cis elements and potential trans-acting factors for the developmental regulation of the Phaseolus vulgaris CHS15 promoter |
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Authors: | Grant S Hotter Jan Kooter Ian A Dubery Christopher J Lamb Richard A Dixon Maria J Harrison |
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Institution: | (1) Plant Biology Division, The Samuel Roberts Noble Foundation, PO Box 2180, 73402 Ardmore, OK, USA;(2) Plant Biology Laboratory, Salk Institute for Biological Studies, 10010 North Torrey Pines Road, 92037 La Jolla, CA, USA;(3) Present address: Molecular Genetics Unit, Department of Microbiology and Genetics, Massey University, Palmerston North, New Zealand;(4) Present address: BioCentrum Amsterdam, IMBW, Dept. Genetics, Vrije Universiteit, De Boelelaan 1087, 1081 HV Amsterdam, Netherlands;(5) Department of Biochemistry, R.A.U. University, Auckland Park, South Africa |
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Abstract: | A nuclear factor (SBF-1) has previously been identified in Phaseolus vulgaris L. (bean) suspension cell nuclear extracts that binds in vitro to three DNase I-footprinted elements (SBF-1 boxes I, II, and III, 5 to 3) in the 5 region of the bean CHS15 (chalcone synthase) gene promoter. To define the functional role of the three SBF-1 boxes in development, we examined transgenic tobacco plants carrying a series of nested CHS15 promoter--glucuronidase (GUS) fusions for GUS activity by histochemical staining. We show that the CHS15 promoter deleted to position-173 and lacking all three SBF-1 boxes directs the same qualitative pattern of expression in initiating lateral roots and in developing seeds as the full length promoter (-326). Thus, activation of expression in these organs is mediated by sequence elements located downstream of the three SBF-1 boxes. However, specific deletions within the-326 to-173 region modulate expression. Thus, deletion of box II abolishes GUS activity in initiating lateral roots. Further deletion of box III fails to restore expression but subsequent deletion of an additional 43 bp to position-173 re-establishes expression. We show that sequence-specific DNA-binding activities consistent with these results are present in nuclear extracts of bean roots and seeds. These studies reveal cis elements within the CHS15 promoter, and potential trans factors, that permit organ- and tissue-specific developmental patterns of regulation to be combined with a flexible response to environmental cues. |
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Keywords: | chalcone synthase cis elements Phaseolus vulgaris seed development |
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