Characterization of the oriT region of the IncFV plasmid pED208.
DNA sequence analysis of a 2.2kb EcoRI-HindIII fragment from pED208, the derepressed form of the IncFV plasmid Folac, revealed sequences highly homologous to the oriT region, traM, and traJ genes of other IncF plasmids. The TraM protein was purified and immunoblots of fractionated cells containing pED208 or Folac showed that TraM was predominantly in the cytoplasm. Using DNA retardation assays and the DNase I footprinting technique, the TraM protein was found to bind to three large motifs in the oriT region: (I) an inverted repeat, (II) two direct repeats, and (III) the traM promoter region. These three footprint regions contained a Hinfl-like sequence (GANTC) that appeared 16 times, spaced 11-12 bp (or multiples thereof) apart, suggesting that TraM protein binds in a complex manner over this entire region.
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Department of Biochemistry, University of Alberta, Edmonton, Canada.
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D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000276:immunology; D001483:Base Sequence; D002458:Cell Fractionation; D003227:Conjugation, Genetic; D004261:DNA Replication / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D005144:F Factor / Q000235:genetics*; D008969:Molecular Sequence Data; D009876:Operon; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C074281:TraM protein, bacterial