title: Profiles of embryonic nuclear protein binding to the proximal promoter region of the soybean β-conglycinin α subunit gene.
abstract: β-Conglycinin, a major component of seed storage protein in soybean, comprises three subunits: α, α' and β. The expression of genes for these subunits is strictly controlled during embryogenesis. The proximal promoter region up to 245 bp upstream of the transcription start site of the α subunit gene sufficiently confers spatial and temporal control of transcription in embryos. Here, the binding profile of nuclear proteins in the proximal promoter region of the α subunit gene was analysed. DNase I footprinting analysis indicated binding of proteins to the RY element and DNA regions including box I, a region conserved in cognate gene promoters. An electrophoretic mobility shift assay (EMSA) using different portions of box I as a probe revealed that multiple portions of box I bind to nuclear proteins. In addition, an EMSA using nuclear proteins extracted from embryos at different developmental stages indicated that the levels of major DNA-protein complexes on box I increased during embryo maturation. These results are consistent with the notion that box I is important for the transcriptional control of seed storage protein genes. Furthermore, the present data suggest that nuclear proteins bind to novel motifs in box I including 5'-TCAATT-3' rather than to predicted cis-regulatory elements.
authors.affiliation: Research Faculty of Agriculture, Hokkaido University, Sapporo, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Yoshino
authors.forename: K
authors.initials: K
authors.lastname: Tsutsumi
authors.forename: A
authors.initials: A
authors.lastname: Kanazawa
mesh_terms: D052179:Antigens, Plant / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D018548:Cotyledon / Q000378:metabolism; D018983:DNA Footprinting; D024202:Electrophoretic Mobility Shift Assay; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D018506:Gene Expression Regulation, Plant; D005916:Globulins / Q000235:genetics* / Q000378:metabolism; D005966:Glucuronidase / Q000378:metabolism; D008969:Molecular Sequence Data; D009687:Nuclear Proteins; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D021122:Protein Subunits; D055314:Seed Storage Proteins / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA; D030262:Soybean Proteins / Q000235:genetics* / Q000378:metabolism; D013025:Glycine max / Q000196:embryology / Q000235:genetics* / Q000378:metabolism
chemical_list: D052179:Antigens, Plant; D005916:Globulins; D009687:Nuclear Proteins; D021122:Protein Subunits; D055314:Seed Storage Proteins; D030262:Soybean Proteins; C013894:beta-conglycinin protein, Glycine max; D005966:Glucuronidase
keywords: DNA-protein interaction; electrophoretic mobility shift assay; embryogenesis; proximal promoter region; seed storage protein; β-conglycinin