Transcriptional regulation of flhDC by QseBC and sigma (FliA) in enterohaemorrhagic Escherichia coli

Mol Microbiol. 2005 Sep;57(6):1734-49. doi: 10.1111/j.1365-2958.2005.04792.x.


Enterohaemorrhagic Escherichia coli (EHEC) serotype O157:H7, the causative agent of haemorrhagic colitis, has been shown to utilize a cell-to-cell signalling system to regulate gene expression. We have previously reported that the quorum sensing E. coli regulators B and C (QseBC) may act as a two-component system in EHEC to transcriptionally regulate the expression of flagella and motility through flhDC, the master regulator of flagella and motility genes. Here, we performed deletion analyses using the flhDC promoter in order to determine the minimal promoter regions necessary for QseBC transcriptional activation. We also performed electrophoretic mobility shift assays, competition experiments and DNaseI footprints, which suggest that QseB directly binds the flhDC promoter at high- and low-affinity binding sites. These analyses have allowed us to determine the potential consensus sequence to which QseB binds in order to regulate transcription. Additionally, we mapped the transcriptional start site of flhDC responsive to QseBC, leading to the identification of a conserved FliA (sigma28) consensus sequence. These results suggest that FliA (sigma28), a class 2 flagellar gene, may be aiding in the transcriptional initiation of class 1 genes (flhDC) in EHEC. In order to further characterize the role of FliA (sigma28) in transcription of the flhDC promoter, we constructed a fliA isogenic mutant in EHEC. The flhDC::lacZ transcriptional fusion showed decreased activity in the fliA mutant compared with wild-type and complemented strains. Taken together, these results indicate that transcriptional initiation at the flhDC promoter by QseBC appears to be complex and dependent on the presence of FliA (sigma28).

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Base Sequence
  • Consensus Sequence
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Escherichia coli O157 / genetics
  • Escherichia coli O157 / growth & development
  • Escherichia coli O157 / metabolism*
  • Escherichia coli O157 / physiology
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism*
  • Flagella / metabolism
  • Gene Expression Regulation, Bacterial*
  • Humans
  • Molecular Sequence Data
  • Promoter Regions, Genetic
  • Sigma Factor / chemistry
  • Sigma Factor / genetics
  • Sigma Factor / metabolism*
  • Trans-Activators / chemistry
  • Trans-Activators / genetics
  • Trans-Activators / metabolism*
  • Transcription, Genetic


  • Bacterial Proteins
  • DNA-Binding Proteins
  • Escherichia coli Proteins
  • FliA protein, Bacteria
  • QseB protein, E coli
  • QseC protein, E coli
  • Sigma Factor
  • Trans-Activators
  • flhC protein, E coli
  • flhD protein, E coli