Abstract
Here we describe a system that enables short peptides to bind DNA sequence-specifically. Linking the peptide covalently to DNA through a disulphide bond eliminates the unfavourable energetic cost of diffusion and thus potentiates the peptide-DNA interaction. By this approach we have deconstructed the GCN4/DNA complex into its elemental DNA recognition units. We find that the GCN4 basic region contacts the two half-sites with very different affinities and propose that this thermodynamic asymmetry plays a role in differential regulation of gene expression. Specific binding of the peptide to DNA stabilizes the disulphide bond toward reduction suggesting a novel approach to the discovery of new DNA-binding specificities.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Base Sequence
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Circular Dichroism
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Consensus Sequence
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DNA / chemistry
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DNA / metabolism*
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DNA-Binding Proteins / chemistry
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DNA-Binding Proteins / metabolism*
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Disulfides / chemistry
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Fungal Proteins / chemistry
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Fungal Proteins / genetics
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Fungal Proteins / metabolism*
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Leucine Zippers / physiology*
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Models, Chemical
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Models, Molecular
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Molecular Sequence Data
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Molecular Structure
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Peptides / chemistry
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Peptides / metabolism*
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Protein Binding
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Protein Kinases / chemistry
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Protein Kinases / genetics
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Protein Kinases / metabolism*
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Saccharomyces cerevisiae Proteins*
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Structure-Activity Relationship
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Sulfhydryl Reagents / chemistry
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Temperature
Substances
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DNA-Binding Proteins
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Disulfides
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Fungal Proteins
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Peptides
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Saccharomyces cerevisiae Proteins
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Sulfhydryl Reagents
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DNA
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Protein Kinases