An element with palindromic structure is required for the expression of TBP (TATA box-binding protein) gene in Drosophila melanogaster.
Previously we showed that the 5'-flanking regions between -261 and -207 of the Drosophila melanogaster TBP (TATA box binding protein) gene is important for its expression. We further made serial deletion mutants in this region and analyzed their promoter activities using the transient transfection assay. We found that the 16 bp deletion from -261 to -245 greatly reduces the promoter activity of the Drosophila TBP gene. The 16 bp DNA element contains half of a 11 bp long palindromic sequence, CTTTT-GAAAAG. Disruption of the palindromic sequence by site-directed mutagenesis severely affected promoter activity. In addition, the electrophoretic mobility shift assay showed that the oligonucleotide containing the palindromic sequence can make specific DNA/protein complexes when it was mixed with the Drosophila nuclear extract, suggesting that it interacts with nuclear protein(s). Our data suggest that the palindromic sequence has a critical role in the expression of the Drosophila TBP gene.
Oh
Y
Y
Department and Institute of Genetic Engineering, Kyung Hee University, Yongin, Korea.
Lee
C
C
Baek
K
K
Kim
W
W
Yoon
J
J
Han
K
K
Cho
N
N
D000818:Animals; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000032:analysis / Q000235:genetics*; D004331:Drosophila melanogaster / Q000235:genetics*; D005786:Gene Expression Regulation / Q000235:genetics; D017344:Genes, Insect; D017930:Genes, Reporter; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D009687:Nuclear Proteins / Q000032:analysis; D011401:Promoter Regions, Genetic / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D035181:TATA-Box Binding Protein; D014157:Transcription Factors / Q000235:genetics*; D014162:Transfection
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D035181:TATA-Box Binding Protein; D014157:Transcription Factors