Abstract
Structure determination of protein-nucleic acid complexes remains a challenging task. Here we present a simple method for generating crystals of a CsrA-nucleic acid complex, guided entirely by results from nuclear magnetic resonances spectroscopy (NMR) spectroscopy. Using a construct that lacks thirteen non-essential C-terminal residues, efficient binding to DNA could be demonstrated. One CsrA dimer interacts with two DNA oligonucleotides, similar to previous findings with RNA. Furthermore, the NMR study of the CsrA-DNA complex was the basis for successfully homing in on conditions that were suitable for obtaining crystals of the CsrA-DNA complex. Our results may be useful for those cases where RNA in protein-nucleic acid complexes may be replaced by DNA.
MeSH terms
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Amino Acid Sequence
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Bacillus subtilis / genetics
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Bacillus subtilis / metabolism*
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Bacterial Proteins / chemistry*
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Bacterial Proteins / metabolism
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Base Sequence
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Binding Sites / genetics
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Crystallography, X-Ray
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DNA / chemistry
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DNA / metabolism
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Escherichia coli Proteins / chemistry
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Escherichia coli Proteins / metabolism
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Molecular Sequence Data
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Nuclear Magnetic Resonance, Biomolecular*
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Nucleic Acid Conformation
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Protein Multimerization / physiology
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RNA / chemistry*
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RNA / metabolism
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RNA-Binding Proteins / chemistry
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RNA-Binding Proteins / metabolism
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Repressor Proteins / chemistry
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Repressor Proteins / metabolism
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Sequence Homology, Amino Acid
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Transcription Factors / chemistry*
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Transcription Factors / metabolism
Substances
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Bacterial Proteins
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CsrA protein, E coli
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Escherichia coli Proteins
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RNA-Binding Proteins
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Repressor Proteins
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Transcription Factors
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RNA
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DNA