Cis-analysis of a seed protein gene promoter: the conservative RY repeat CATGCATG within the legumin box is essential for tissue-specific expression of a legumin gene.
A 2.4 kb fragment containing the 5'-flanking region and the 5'-noncoding sequence of the Vicia faba legumin gene LeB4 mediates high level seed-specific expression in transgenic tobacco plants. Deleted derivatives of this legumin upstream sequence were fused to the npt-II reporter gene to determine the tissue-specific activity of the chimeric constructs in stably transformed tobacco plants. The results indicate the presence of positive regulatory, enhancer-like cis elements within 566 bp of the upstream sequence. Most importantly, however, these elements are only fully functional in conjunction with the core motif CATGCATG of the legumin box around position -95, since destruction of the motif by a 6 bp deletion in an otherwise intact 2.4 kb upstream sequence drastically reduces expression in seeds. At the same time, low level expression in leaves is observed. The occurrence of similar CATGCATG consensus cis elements with alternating purine and pyrimidine base pairs in front of several other plant genes suggests a functional role of the motif in a wider range of plant promoters.
Bäumlein
H
H
Institut für Genetik und Kulturpflanzenforschung, Gatersleben, Germany.
Nagy
I
I
Villarroel
R
R
Inzé
D
D
Wobus
U
U
D001483:Base Sequence; D017124:Conserved Sequence; D004252:DNA Mutational Analysis; D007887:Fabaceae / Q000235:genetics*; D019868:Kanamycin Kinase; D008969:Molecular Sequence Data; D010770:Phosphotransferases / Q000096:biosynthesis / Q000235:genetics; D010940:Plant Proteins; D014674:Plant Proteins, Dietary / Q000235:genetics*; D010946:Plants, Medicinal; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012091:Repetitive Sequences, Nucleic Acid; D012639:Seeds / Q000378:metabolism; D017384:Sequence Deletion; D012689:Sequence Homology, Nucleic Acid; D014018:Tissue Distribution; D014158:Transcription, Genetic; D014170:Transformation, Genetic; D000097793:Legumins
D019868:Kanamycin Kinase; D010770:Phosphotransferases; D010940:Plant Proteins; D014674:Plant Proteins, Dietary; D011993:Recombinant Fusion Proteins; D000097793:Legumins