In vitro transcription from the Nectria haematococca PDA1 promoter in an homologous extract reflects in vivo pisatin-responsive regulation.
The PDA1 gene of Nectria haematococca MP VI (anamorph: Fusarium solani) encodes pisatin demethylase. This enzyme detoxifies the isoflavanoid phytoalexin pisatin produced by the plant on which this fungus is pathogenic. Expression of pisatin demethylase activity is induced in a mycelium by pretreatment with pisatin. We have developed homologous in vitro system which accurately initiates transcription from the PDA1 promoter. Transcription levels in vitro reflect the same pisatin-responsive stimulation as measured for PDA1 mRNA in vivo, and are dependent upon sequences in the 5' upstream region of PDA1. Pisatin-responsive transcription from the PDA1 promoter indicates that initiation of transcription is a major regulatory step in the pisatin induction of pisatin demethylase expression.
Ruan
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Department of Botany, University of Maryland, College Park 20742, USA.
Straney
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D001483:Base Sequence; D001578:Benzopyrans / Q000494:pharmacology*; D002474:Cell-Free System; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000235:genetics*; D004790:Enzyme Induction / Q000187:drug effects; D005656:Fungal Proteins / Q000378:metabolism; D005670:Fusarium / Q000201:enzymology* / Q000235:genetics; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects*; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D010090:Oxidoreductases, O-Demethylating / Q000096:biosynthesis / Q000235:genetics*; D018532:Pisum sativum / Q000378:metabolism / Q000382:microbiology; D011401:Promoter Regions, Genetic; D036343:Pterocarpans; D012319:RNA Polymerase II / Q000378:metabolism; D017384:Sequence Deletion; D016385:TATA Box; D014158:Transcription, Genetic; D000094162:Phytoalexins
D001578:Benzopyrans; D003577:Cytochrome P-450 Enzyme System; D005656:Fungal Proteins; D010090:Oxidoreductases, O-Demethylating; D036343:Pterocarpans; D012319:RNA Polymerase II; C057976:pisatin demethylase; C003161:pisatin; D000094162:Phytoalexins