Structural and functional characterization of the phytoene synthase promoter from Arabidopsis thaliana.
The expression of the gene coding for the carotenogenic enzyme phytoene synthase is highly regulated. To study this, its promoter and truncated versions thereof were translationally fused to the luciferase gene as a reporter and these constructs were used to transform Arabidopsis thaliana. The full-length promoter was shown to be active in the dark, but mediated positive responses towards different light qualities (far-red, red, blue and white light). Among the herbicides tested, norflurazon and gabaculine showed no notable effects, while CPTA abolished light induction completely. Response towards different light qualities was mediated by a TATA box-proximal promoter region up to position -300, containing G-box-like elements involved in the distinction of different monochromatic light qualities applied. This is detected in electrophoretic mobility shift assays (EMSAs), which reveal differential complex formation. A TATA box distal region of the promoter was shown to be responsible for a high basal promoter activity that was not modulated by different light qualities. Using EMSAs, a novel cis-acting element ATCTA occurring in tandem between positions -854 and -841 proved to be decisive in this respect. The motif was found in several other promoter regions involved in carotenoid and tocopherol biosynthesis, as well as in the promoter regions mediating the expression of photosynthesis-related genes. The functional equivalence of the motifs was shown by successfully using the respective regions in EMSAs. We conclude that the ATCTA motif represents an element capable of mediating a coordinated regulation of these pathways at the transcriptional level.
Welsch
Ralf
R
Center for Applied Biosciences, Universität Freiburg, Schänzlestrasse 1, 79104 Freiburg, Germany.
Medina
Joaquin
J
Giuliano
Giovanni
G
Beyer
Peter
P
Von Lintig
Johannes
J
D019883:Alkyl and Aryl Transferases / Q000235:genetics*; D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D001483:Base Sequence; D002338:Carotenoids / Q000037:antagonists & inhibitors / Q000378:metabolism; D002734:Chlorophyll / Q000037:antagonists & inhibitors / Q000378:metabolism; D003001:Cloning, Molecular; D024202:Electrophoretic Mobility Shift Assay; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects / Q000528:radiation effects; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000528:radiation effects; D018506:Gene Expression Regulation, Plant / Q000187:drug effects / Q000528:radiation effects; D051232:Geranylgeranyl-Diphosphate Geranylgeranyltransferase; D006540:Herbicides / Q000494:pharmacology; D008027:Light; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009024:Morphogenesis / Q000235:genetics; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D016385:TATA Box / Q000235:genetics
D006540:Herbicides; D002734:Chlorophyll; D002338:Carotenoids; D008156:Luciferases; D019883:Alkyl and Aryl Transferases; D051232:Geranylgeranyl-Diphosphate Geranylgeranyltransferase