Abstract
The Thyroid Transcription factor 1 (TTF-1) recognizes specific DNA sequences by a Homeodomain (TTF-1 HD). The TTF-1 HD DNA-binding properties with both specific and non-specific DNA sequences were investigated. TTF-1 HD exists as a monomer in solution and as a monomer binds DNA. At 75 mM KCl, its relative binding affinity with a specific DNA sequence is about 50 fold higher than with a non-specific DNA sequence. Increase of KCl concentration reduces the apparent binding affinity both to specific and non-specific DNA sequences. However, non-specific binding is more sensitive than specific binding to the increase of salt concentration. When DNA-binding reactions are performed at temperature and salt concentration close to the intracellular environment, TTF-1 HD binds the specific sequence with an affinity at least 1000 fold higher respect to the non-specific sequence.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Base Sequence
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Binding Sites
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Cattle
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Chromatography, Gel
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Cloning, Molecular
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DNA / metabolism*
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DNA-Binding Proteins / metabolism*
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Kinetics
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Molecular Sequence Data
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Nuclear Proteins / biosynthesis
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Nuclear Proteins / isolation & purification
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Nuclear Proteins / metabolism*
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Oligodeoxyribonucleotides
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Osmolar Concentration
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Potassium Chloride / pharmacology
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Promoter Regions, Genetic
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Thermodynamics
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Thyroid Nuclear Factor 1
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Transcription Factors / biosynthesis
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Transcription Factors / isolation & purification
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Transcription Factors / metabolism*
Substances
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DNA-Binding Proteins
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Nuclear Proteins
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Oligodeoxyribonucleotides
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Recombinant Proteins
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Thyroid Nuclear Factor 1
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Transcription Factors
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Potassium Chloride
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DNA