The promoter of the strictosidine synthase gene from periwinkle confers elicitor-inducible expression in transgenic tobacco and binds nuclear factors GT-1 and GBF.
Strictosidine synthase (STR) is a key enzyme in the biosynthesis of terpenoid indole alkaloids. This class of secondary metabolites harbours several pharmaceutically important compounds used, among other applications, in cancer treatment. Terpenoid indole alkaloid biosynthesis and expression of biosynthetic genes including Str1 is induced by fungal elicitors. To identify elicitor-responsive regulatory promoter elements and trans-acting factors, the single-copy Str1 gene was isolated from the subtropical plant species Catharanthus roseus (Madagascar periwinkle). Str1 upstream sequences conferred elicitor-responsive expression to the beta-glucuronidase (gusA) reporter gene in transgenic tobacco plants. Main enhancer sequences within the Str1 promoter region studied were shown to be located between -339 and -145. This region and two other regions of the promoter bound the tobacco nuclear protein factor GT-1. A G-box located around position -105 bound nuclear and cloned G-box-binding factors (GBFs). A mutation that knocked out GBF binding had no measurable effect on expression, which indicates that the G-box is not essential for the elicitor responsiveness of the Str1 promoter. No obvious homologies with promoter elements identified in other elicitor-responsive genes were observed, suggesting that the Str1 gene may depend on novel regulatory mechanisms.
Pasquali
G
G
Institute of Molecular Plant Sciences, Clusius Laboratory, Leiden, Netherlands.
Erven
A S
AS
Ouwerkerk
P B
PB
Menke
F L
FL
Memelink
J
J
D001483:Base Sequence; D019759:Carbon-Nitrogen Lyases / Q000235:genetics*; D002457:Cell Extracts / Q000494:pharmacology; D003001:Cloning, Molecular; D018983:DNA Footprinting; D015342:DNA Probes; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004742:Enhancer Elements, Genetic / Q000235:genetics; D051790:G-Box Binding Factors; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D017343:Genes, Plant / Q000235:genetics; D017930:Genes, Reporter; D019684:Magnoliopsida / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism*; D018515:Plant Leaves / Q000187:drug effects / Q000235:genetics; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D020218:Response Elements / Q000235:genetics; D017384:Sequence Deletion; D014026:Nicotiana / Q000187:drug effects / Q000235:genetics; D015534:Trans-Activators / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*
D002457:Cell Extracts; D015342:DNA Probes; D004268:DNA-Binding Proteins; D051790:G-Box Binding Factors; C077926:GT-1 transcription factor; D009687:Nuclear Proteins; D012333:RNA, Messenger; D015534:Trans-Activators; D014157:Transcription Factors; D019759:Carbon-Nitrogen Lyases; C022684:strictosidine synthetase