Analysis of nuclear proteins interacting with a wheat alpha/beta-gliadin seed storage protein gene.
The promoter region (-524 to -46) of the wheat alpha/beta-gliadin seed storage protein gene was analyzed for interactions with nuclear proteins from developing wheat seeds. Six complexes were detected within the first 165 bp upstream of the transcriptional start site. One of the proteins was a non-sequence specific AT-binding protein. The remaining five proteins bound in a sequence specific manner. One (CABP) mapped to a conserved CA-rich element at -134 to -112 while another (PalBP) mapped to an adjacent, palindromic sequence at -112 to -106. Three proteins (CTBPs 1-3) formed complexes at two, independent homologous sites. The activities of four of the binding proteins, CTBPs 1-3 and CABP, exhibited similar patterns of expression during seed development: they first appeared at early to mid stages, reached a maximum at mid stage and subsequently decreased, paralleling the pattern of gliadin mRNA accumulation. The non-specific AT-binding protein was detected at relatively high levels only at mid development. PalBP activity, on the other hand, first appeared at mid stage and was present at a constant level throughout later stages of development. The results suggest that the binding proteins may regulate gliadin expression in an antagonistic manner.
Institute of Biological Chemistry, Washington State University, Pullman 99164-6340.
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Vellanoweth
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Okita
D001483:Base Sequence; D017124:Conserved Sequence; D015342:DNA Probes; D004268:DNA-Binding Proteins / Q000032:analysis; D004742:Enhancer Elements, Genetic / Q000235:genetics; D005786:Gene Expression Regulation; D017343:Genes, Plant / Q000235:genetics*; D005903:Gliadin / Q000235:genetics*; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000032:analysis*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D012639:Seeds / Q000737:chemistry; D013997:Time Factors; D014908:Triticum / Q000737:chemistry / Q000196:embryology*
D015342:DNA Probes; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D012333:RNA, Messenger; D005903:Gliadin