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Identification of elements in thePDA1 promoter ofNectria haematococca necessary for a high level of transcription in vitro

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Abstract

Expression of thePDA1 gene in the ascomyceteNectria haematococca MPVI (anamorph:Fusarium solani) is induced by exposure of mycelium to pisatin, an isoflavonoid phytoalexin produced by its host plant, garden pea. ThePDA1 gene encodes a cytochrome P-450 monooxygenase which detoxifies pisatin. Regulatory elements controlling transcription from thePDA1 promoter were identified using a homologousNectria in vitro transcription system through analysis of 5′ deletions, specific oligonucleotide competition, and fusion of upstream segments to a heterologous promoter. A promoter-distal element which provided transcriptional activation was localized to a 35-bp region positioned − 514 to − 483 upstream of the transcriptional start site. This 35-bp region binds a previously characterized pisatin-responsive DNA-binding factor (PRF) and thus may provide pisatin-responsive control of transcription. A second promoter-proximal positive-acting region was found to be necessary for promoter transcription in both homologous and heterologous extracts, and so is likely to bind less gene-specific transcription activator(s). A negative-acting element located between these two positive regions may act to make the positive-acting elements interdependent. The identification of an activator responding to pisatin provides a model for the control of a number of genes and processes controlled by host-specific signals, particularly the flavonoids.

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Author notes

  1. Yijun Ruan

    Present address: Plant Genome Expression Program, U3D, Monsanto Co., 800 N. Lindberg Blvd., 63167, St Louis, MO, USA

Authors and Affiliations

  1. Department of Plant Biology and Maryland Agriculture Experiment Station, University of Maryland, 20742, College Park, MD, USA

    Yijun Ruan & David C. Straney

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  1. Yijun Ruan
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  2. David C. Straney
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Communicated by C. van den Hondel

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Ruan, Y., Straney, D.C. Identification of elements in thePDA1 promoter ofNectria haematococca necessary for a high level of transcription in vitro. Molec. Gen. Genet. 250, 29–38 (1996). https://doi.org/10.1007/BF02191822

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  • DOI: https://doi.org/10.1007/BF02191822

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