The Aspergillus nidulans transcription factor AlcR forms a stable complex with its half-site DNA: a NMR study.
The Aspergillus nidulans transcription factor AlcR is shown by NMR and gel retardation assay to form a stable complex with oligonucleotide sequences comprising the consensus half-site 5'-TGCGG-3'. Apparent microM dissociation constants are evaluated by both methods. The measured lifetime of the complex is 74+/-7 ms at 20 degrees C with the following DNA sequence: 5'-C1G2T3G4C5G6G7A8T9C10-3'. The major chemical shift variations upon binding involve both the two adjacent GC pairs (G6 and G7) and, clearly, the AT pairs at both ends of the consensus sequence (T3 and A8), suggesting additional contacts of the protein with the DNA. This extensive and strong interaction with the half-site is another example of the variability in contacts of the fungal DNA-binding proteins containing Zn2Cys6 domains with their consensus sites. It is the first demonstration that a binuclear cluster protein can bind to DNA as a monomer with strong affinity.
Cerdan
R
R
Laboratoire de RMN, ICSN-CNRS, Gif-sur-Yvette, France.
Collin
D
D
Lenouvel
F
F
Felenbok
B
B
Guittet
E
E
D001233:Aspergillus nidulans / Q000737:chemistry* / Q000378:metabolism; D001665:Binding Sites; D016384:Consensus Sequence; D004271:DNA, Fungal / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*; D015032:Zinc / Q000737:chemistry / Q000378:metabolism
C074202:ALCR protein, Aspergillus nidulans; D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D009838:Oligodeoxyribonucleotides; D014157:Transcription Factors; D015032:Zinc