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Profiles of embryonic nuclear protein binding to the proximal promoter region of the soybean β‐conglycinin α subunit gene
Author(s) -
Yoshino M.,
Tsutsumi K.,
Kanazawa A.
Publication year - 2015
Publication title -
plant biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.871
H-Index - 87
eISSN - 1438-8677
pISSN - 1435-8603
DOI - 10.1111/plb.12218
Subject(s) - biology , electrophoretic mobility shift assay , nuclear protein , promoter , gene , transcription factor , transcription (linguistics) , protein subunit , microbiology and biotechnology , caat box , footprinting , dna binding protein , gene expression , genetics , linguistics , philosophy
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. DN ase 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.