Sequence-specific interactions of a nuclear protein factor with the promoter region of a rice gene for alpha-amylase, RAmy3D.
The expression of a rice gene for alpha-amylase, RAmy3D, in suspension-cultured cells is induced at the transcriptional level by the deprivation of sugars. Binding of a nuclear protein from suspension-cultured rice cells to the promoter region of the RAmy3D gene was studied by gel-retardation and DNase I footprinting assays. Gel-retardation assays indicated that a 358-bp fragment of the promoter region interacted specifically with a protein factor from suspension-cultured cells. DNase I footprinting analysis allowed us to define three protein-binding regions. Each of these protein-binding sequences contained the GCCG G/C CG motif, which is specifically present in the promoter region of the sugar-regulated gene, RAmy3D, for rice alpha-amylase and not in that of the gibberellin-regulated RAmy1A gene. Subsequent cross-competition experiments using gel-retardation assay and synthetic oligonucleotides showed that the GCCG G/C CG motifs directly mediated the binding of a nuclear protein. These observations are discussed in relation to expression of the gene for alpha-amylase in suspension-cultured cells.
Nagoya University, BioScience Center, Japan.
S
S
Mitsunaga
R L
RL
Rodriguez
J
J
Yamaguchi
D001483:Base Sequence; D050260:Carbohydrate Metabolism; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D003850:Deoxyribonuclease I; D005786:Gene Expression Regulation; D017343:Genes, Plant; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism*; D012275:Oryza / Q000166:cytology / Q000201:enzymology / Q000235:genetics*; D010940:Plant Proteins / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D000516:alpha-Amylases / Q000235:genetics*
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D010940:Plant Proteins; D004247:DNA; D003850:Deoxyribonuclease I; D000516:alpha-Amylases