Detection of DNA bending in a DNA-PAP1 protein complex by fluorescence resonance energy transfer.
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.
Ozaki
H
H
Department of Chemistry, Gunma University, Japan.
Iwase
N
N
Sawai
H
H
Kodama
T
T
Kyogoku
Y
Y
D050976:Basic-Leucine Zipper Transcription Factors; D001665:Binding Sites; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D003852:Deoxyribonucleoproteins / Q000737:chemistry*; D004735:Energy Transfer; D005656:Fungal Proteins / Q000737:chemistry; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D000073718:Pancreatitis-Associated Proteins; D012568:Schizosaccharomyces; D029702:Schizosaccharomyces pombe Proteins; D013050:Spectrometry, Fluorescence; D014157:Transcription Factors / Q000737:chemistry*
D050976:Basic-Leucine Zipper Transcription Factors; D004268:DNA-Binding Proteins; D003852:Deoxyribonucleoproteins; D005656:Fungal Proteins; D009838:Oligodeoxyribonucleotides; D000073718:Pancreatitis-Associated Proteins; C105096:Pap1 protein, S pombe; C000612141:REG3A protein, human; D029702:Schizosaccharomyces pombe Proteins; D014157:Transcription Factors