Characterization of a new LexA binding motif in the marine magnetotactic bacterium strain MC-1

J Bacteriol. 2003 Aug;185(15):4471-82. doi: 10.1128/JB.185.15.4471-4482.2003.

Abstract

MC-1 is a marine, magnetotactic bacterium that is phylogenetically associated with the alpha subclass of the Proteobacteria and is the first and only magnetotactic coccus isolated in pure culture to date. By using a TBLASTN search, a lexA gene was identified in the published genome of MC-1; it was subsequently cloned, and the protein was purified to >90% purity. Results from reverse transcription-PCR analysis revealed that the MC-1 lexA gene comprises a single transcriptional unit with two open reading frames encoding proteins of unknown function and with a rumA-like gene, a homologue of the Escherichia coli umuD gene. Mobility shift assays revealed that this LexA protein specifically binds both to its own promoter and to that of the umuDC operon. However, MC-1 LexA does not bind to the promoter regions of other genes, such as recA and uvrA, that have been previously reported to be regulated by LexA in bacterial species belonging to the alpha subclass of the Proteobacteria: Site-directed mutagenesis of both the lexA and umuDC operator regions demonstrated that the sequence CCTN(10)AGG is the specific target motif for the MC-1 LexA protein.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Alphaproteobacteria / metabolism*
  • Amino Acid Motifs
  • Amino Acid Sequence
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Base Sequence
  • Binding Sites
  • DNA-Directed DNA Polymerase
  • Electrophoretic Mobility Shift Assay
  • Escherichia coli Proteins / genetics*
  • Escherichia coli Proteins / metabolism
  • LexA Repressor Protein
  • Magnetics*
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Operon
  • Promoter Regions, Genetic*
  • Regulon
  • Seawater / microbiology*
  • Sequence Analysis, DNA
  • Serine Endopeptidases / genetics
  • Serine Endopeptidases / metabolism*

Substances

  • Bacterial Proteins
  • DNA-Directed DNA Polymerase
  • Escherichia coli Proteins
  • Serine Endopeptidases
  • LexA Repressor Protein
  • UmuC protein, E coli
  • UmuD protein, E coli