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KMID : 0624620140470020092
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BMB Reports
2014 Volume.47 No. 2 p.92 ~ p.97
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Sequence-specific interaction between ABD-B homeodomain and castor gene in Drosophila
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Kim Keon-Hee
Yoo Si-Uk
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Abstract
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We have examined the effect of bithorax complex genes on the expression of castor gene. During the embryonic stages 12-15, both Ultrabithorax and abdominal-A regulated the castor gene expression negatively, whereas Abdominal-B showed a positive correlation with the castor gene expression according to real-time PCR. To investigate whether ABD-B protein directly interacts with the castor gene, electrophoretic mobility shift assays were performed using the recombinant ABD-B homeodomain and oligonucleotides, which are located within the region 10 kb upstream of the castor gene. The results show that ABD-B protein directly binds to the castor gene specifically. ABD-B binds more strongly to oligonucleotides containing two 5'-TTAT-3' canonical core motifs than the probe containing the 5'-TTAC-3' motif. In addition, the sequences flanking the core motif are also involved in the protein-DNA interaction. The results demonstrate the importance of HD for direct binding to target sequences to regulate the expression level of the target genes.
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KEYWORD
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Abdominal-B, Castor, EMSA, Homeodomain, Homeotic gene
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