Abstract
NHP6A is a non-sequence-specific DNA-binding protein from Saccharomyces cerevisiae which belongs to the HMGB protein family. Previously, we have solved the structure of NHP6A in the absence of DNA and modeled its interaction with DNA. Here, we present the refined solution structures of the NHP6A-DNA complex as well as the free 15bp DNA. Both the free and bound forms of the protein adopt the typical L-shaped HMGB domain fold. The DNA in the complex undergoes significant structural rearrangement from its free form while the protein shows smaller but significant conformational changes in the complex. Structural and mutational analysis as well as comparison of the complex with the free DNA provides insight into the factors that contribute to binding site selection and DNA deformations in the complex. Further insight into the amino acid determinants of DNA binding by HMGB domain proteins is given by a correlation study of NHP6A and 32 other HMGB domains belonging to both the DNA-sequence-specific and non-sequence-specific families of HMGB proteins. The resulting correlations can be rationalized by comparison of solved structures of HMGB proteins.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid 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 / genetics
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DNA-Binding Proteins / metabolism
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HMGN Proteins
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Humans
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Hydrogen Bonding
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Lymphoid Enhancer-Binding Factor 1
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Macromolecular Substances
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Magnetic Resonance Spectroscopy
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Methionine / chemistry
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Models, Molecular
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Molecular Sequence Data
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Molecular Structure
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Nuclear Proteins / chemistry*
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Nuclear Proteins / genetics
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Nuclear Proteins / metabolism
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Nucleic Acid Conformation*
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Phenylalanine / chemistry
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Phylogeny
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Protein Binding
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Protein Structure, Tertiary*
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Repressor Proteins / chemistry
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Repressor Proteins / genetics
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Repressor Proteins / metabolism
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Saccharomyces cerevisiae / chemistry*
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Saccharomyces cerevisiae / genetics
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Saccharomyces cerevisiae / metabolism
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Saccharomyces cerevisiae Proteins / chemistry*
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / metabolism
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Sequence Alignment
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Sex-Determining Region Y Protein
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Transcription Factors / chemistry
Substances
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DNA-Binding Proteins
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HMGN Proteins
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Lymphoid Enhancer-Binding Factor 1
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Macromolecular Substances
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NHP6A protein, S cerevisiae
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Nuclear Proteins
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Repressor Proteins
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Saccharomyces cerevisiae Proteins
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Sex-Determining Region Y Protein
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Transcription Factors
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Phenylalanine
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DNA
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Methionine