Conversion of GC-->AT recognition and elucidation of AT recognition mechanism in zinc finger transcription factor by permutational approach.
Zinc finger-DNA recognition has been investigated through GC-->AT conversion of recognition sequence 5'-GGG-GCG-GGG-3' of transcription factor Sp1 containing three zinc fingers. Three critical amino acid residues of the finger 2 in Sp1 were replaced with the corresponding residues of the finger 4 in CF2-II. The finger 2 of the hybrid protein is expected to recognize 5'-ATA-3' sequence. Indeed, the designed protein Sp1QTQ2 specifically bound to ATA and ATAN probes. The present study demonstrates that one zinc finger recognizes total sequence of the target DNA site rather than triplet base sequence.
Emori
T
T
Institute for Chemical Research, Kyoto University, Japan.
Nagaoka
M
M
Sugiura
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics* / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D029721:Drosophila Proteins; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D066298:In Vitro Techniques; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016329:Sp1 Transcription Factor / Q000737:chemistry / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016335:Zinc Fingers / Q000235:genetics / Q000502:physiology*
D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D011994:Recombinant Proteins; D016329:Sp1 Transcription Factor; D014157:Transcription Factors; C076904:Cf2 protein, Drosophila; D004247:DNA