DNA binding by Corynebacterium glutamicum TetR-type transcription regulator AmtR

BMC Mol Biol. 2009 Jul 23;10:73. doi: 10.1186/1471-2199-10-73.

Abstract

Background: The TetR family member AmtR is the central regulator of nitrogen starvation response in Corynebacterium glutamicum. While the AmtR regulon was physiologically characterized in great detail up to now, mechanistic questions of AmtR binding were not addressed. This study presents a characterization of functionally important amino acids in the DNA binding domain of AmtR and of crucial nucleotides in the AmtR recognition motif.

Results: Site-directed mutagenesis, the characterization of corresponding mutant proteins by gel retardation assays and surface plasmon resonance and molecular modelling revealed several amino acids, which are directly involved in DNA binding, while others have more structural function. Furthermore, we could show that the spacing of the binding motif half sites is crucial for repression of transcription by AmtR.

Conclusion: Although the DNA binding domain of TetR-type repressors is highly conserved and a core binding motif was identified for AmtR and TetR(D), the AmtR binding domain shows individual properties compared to other TetR proteins. Besides by distinct amino acids of AmtR, DNA binding is influenced by nucleotides not only of the conserved binding motif but also by spacing nucleotides in C. glutamicum.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Base Sequence
  • Binding Sites
  • Conserved Sequence
  • Corynebacterium glutamicum / metabolism*
  • DNA, Bacterial / metabolism*
  • Gene Expression Regulation, Bacterial*
  • Models, Molecular
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • Protein Binding
  • Protein Structure, Tertiary
  • Repressor Proteins / chemistry
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism*
  • Sequence Alignment
  • Trans-Activators / metabolism
  • Transcription, Genetic*

Substances

  • AmtR protein, Corynebacterium glutamicum
  • Bacterial Proteins
  • DNA, Bacterial
  • Repressor Proteins
  • TetR protein, Clostridium tetani
  • Trans-Activators