A strong constitutive positive element is essential for the ammonium-regulated expression of a soybean gene encoding cytosolic glutamine synthetase

In order to identify important promoter elements controlling the ammonium-regulated expression of the soybean gene GS15 encoding cytosolic glutamine synthetase, a series of 5' promoter deletions were fused to the GUS reporter gene. To allow the detection of positive and negative regulatory elem...

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Bibliographic Details
Published in:Plant molecular biology 1999-02, Vol.39 (3), p.551-564
Main Authors: Terce-Laforgue, T. (Laboratoire du Metabolisme et de la Nutrition des Plantes INRA, Versailles (France).), Carrayol, E, Cren, M, Desbrosses, G, Hecht, V, Hirel, B
Format: Article
Language:English
Subjects:
ACTIVADOR GENICO
ACTIVATEUR GENIQUE
Agrobacterium rhizogenes
AMMONIA
AMMONIAC
AMONIACO
Bacteria
Base Sequence
Cytosol - enzymology
DNA, Plant - chemistry
DNA, Plant - genetics
EXPRESION GENICA
EXPRESSION DES GENES
Fabaceae - enzymology
Fabaceae - genetics
GENE ACTIVATORS
GENE EXPRESSION
Gene Expression - drug effects
Gene Expression Regulation, Enzymologic
Gene Expression Regulation, Plant
Genes, Plant - genetics
GLUTAMATE AMMONIA LIGASE
Glutamate-Ammonia Ligase - genetics
Glutamate-Ammonia Ligase - metabolism
GLUTAMINA SINTETASA
GLUTAMINE SYNTHETASE
GLYCINE MAX
Glycine max - chemistry
Glycine max - enzymology
Glycine max - genetics
LOTUS CORNICULATUS
Molecular Sequence Data
NODOSITE RACINAIRE
NUCLEOTIDE SEQUENCE
NUDOSIDADES RADICULARES
PLANTAS TRANSGENICAS
PLANTE TRANSGENIQUE
Plants, Genetically Modified
Plants, Medicinal
Promoter Regions, Genetic
Quaternary Ammonium Compounds - pharmacology
Regulatory Sequences, Nucleic Acid
ROOT NODULES
SECUENCIA NUCLEOTIDICA
Sequence Alignment
Sequence Analysis, DNA
Sequence Deletion
SEQUENCE NUCLEOTIDIQUE
TRANSGENIC PLANTS
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Summary:In order to identify important promoter elements controlling the ammonium-regulated expression of the soybean gene GS15 encoding cytosolic glutamine synthetase, a series of 5' promoter deletions were fused to the GUS reporter gene. To allow the detection of positive and negative regulatory elements, a series of 3' deletions were fused to a -90 CaMV 35S promoter fragment placed upstream of the GUS gene. Both types of construct were introduced into Lotus corniculatus plants and soybean roots via Agrobacterium rhizogenes-mediated transformation. Both spectrophotometric enzymatic analysis and histochemical localization of GUS activity in roots, root nodules and shoots of transgenic plants revealed that a strong constitutive positive element (SCPE) of 400 bp, located in the promoter distal region is indispensable for the ammonium-regulated expression of GS15. Interestingly, this SCPE was able to direct constitutive expression in both a legume and non-legume background to a level similar to that driven by the CaMV 35S full-length promoter. In addition, results showed that separate proximal elements, located in the first 727 bp relative to the transcription start site, are essential for root- and root nodule-specific expression. This proximal region contains an AAAGAT and two TATTTAT consensus sequences characteristic of nodulin or nodule-enhanced gene promoters. A putative silencer region containing the same TATTTAT consensus sequence was identified between the SCPE and the organ-specific elements. The presence of positive, negative and organ-specific elements together with the three TATTTAT consensus sequences within the promoter strongly suggest that these multiple promoter fragments act in a cooperative manner, depending on the spatial conformation of the DNA for trans-acting factor accessibility.
ISSN:0167-4412
1573-5028
DOI:10.1023/A:1006169018296