Abstract
The structure of DNA in a DNA-protein complex was studied by means of fluorescence resonance energy transfer (FRET) method. Oligonucleotide phosphorothioates were labeled with fluorescein and eosin to obtain a donor- and acceptor-labeled DNA. The formation of a complex of the DNA with PAP1(70), which is a DNA binding site fragment derived from transcription regulatory protein, PAP1, of fission yeast, was confirmed by gel retardation analysis and fluorescence measurements. FRET of the donor- and acceptor-labeled DNA with and without PAP1(70) indicated that the DNA in the complex was bent about 26 degrees toward the protein-binding surface.
MeSH terms
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Basic-Leucine Zipper Transcription Factors
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Binding Sites
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DNA-Binding Proteins / chemistry*
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DNA-Binding Proteins / metabolism
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Deoxyribonucleoproteins / chemistry*
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Energy Transfer
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Fungal Proteins / chemistry
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Nucleic Acid Conformation*
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Oligodeoxyribonucleotides / chemistry
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Pancreatitis-Associated Proteins
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Schizosaccharomyces
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Schizosaccharomyces pombe Proteins
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Spectrometry, Fluorescence
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Transcription Factors / chemistry*
Substances
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Basic-Leucine Zipper Transcription Factors
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DNA-Binding Proteins
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Deoxyribonucleoproteins
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Fungal Proteins
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Oligodeoxyribonucleotides
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Pancreatitis-Associated Proteins
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Pap1 protein, S pombe
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REG3A protein, human
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Schizosaccharomyces pombe Proteins
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Transcription Factors