DNA-binding properties of Mblk-1, a putative transcription factor from the honeybee.
We previously identified a gene, Mblk-1, that encodes a putative transcription factor with two DNA-binding motifs expressed preferentially in the honeybee brain [H. Takeuchi et al., Insect Mol. Biol. 10, 487-494 (2001)]. In the present study, we identified its preferred binding sequence as 5'-CCCTATCGATCGATCTCTACCT-3' and characterized its DNA-binding properties using truncated Mblk-1 mutants. An electrophoretic mobility shift assay revealed that the full-length Mblk-1 binds to the sequence with high affinity, whereas each truncated DNA-binding motif of Mblk-1 binds with much lower affinities. An in vitro pull-down assay indicated that each DNA-binding motif affords homodimeric bindings, suggesting that Mblk-1 functions as a dimer.
Park
Jung-Min
JM
Department of Biological Sciences, Graduate School of Science, University of Tokyo, Bunkyo-ku, Tokyo, 113-0033, Japan.
Kunieda
Takekazu
T
Takeuchi
Hideaki
H
Kubo
Takeo
T
D020816:Amino Acid Motifs / Q000502:physiology; D000818:Animals; D001516:Bees; D001667:Binding, Competitive / Q000502:physiology; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D019281:Dimerization; D016297:Mutagenesis, Site-Directed; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000502:physiology*; D017422:Sequence Analysis, DNA; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
D004268:DNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D004247:DNA