PMID:21768111

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Citation

McVicker, G, Sun, L, Sohanpal, BK, Gashi, K, Williamson, RA, Plumbridge, J and Blomfield, IC (2011) SlyA protein activates fimB gene expression and type 1 fimbriation in Escherichia coli K-12.J. Biol. Chem. 286:32026-35

Abstract

We have demonstrated that SlyA activates fimB expression and hence type 1 fimbriation, a virulence factor in Escherichia coli. SlyA is shown to bind to two operator sites (O(SA1) and O(SA2)), situated between 194 and 167 base pairs upstream of the fimB transcriptional start site. fimB expression is derepressed in an hns mutant and diminished by a slyA mutation in the presence of H-NS only. H-NS binds to multiple sites in the promoter region, including two sites (H-NS2 and H-NS3) that overlap O(SA1) and O(SA2), respectively. Mutations that disrupt either O(SA1) or O(SA2) eliminate or reduce the activating effect of SlyA but have different effects on the level of expression. We interpret these results as reflecting the relative competition between SlyA and H-NS binding. Moreover we show that SlyA is capable of displacing H-NS from its binding sites in vitro. We suggest SlyA binding prevents H-NS binding to H-NS2 and H-NS3 and the subsequent oligomerization of H-NS necessary for full inhibition of fimB expression. In addition, we show that SlyA activates fimB expression independently of two other known regulators of fimB expression, NanR and NagC. It is demonstrated that the rarely used UUG initiation codon limits slyA expression and that low SlyA levels limit fimB expression. Furthermore, Western blot analysis shows that cells grown in rich-defined medium contain ~1000 SlyA dimers per cell whereas those grown in minimal medium contain >20% more SlyA. This study extends our understanding of the role that SlyA plays in the host-bacterial relationship.

Links

PubMed Online version:10.1074/jbc.M111.266619

Keywords


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Materials and Methods Used

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Phenotype Annotations

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Phenotype of Taxon Information Genotype Information (if known) Condition Information OMP ID OMP Term Name ECO ID ECO Term Name Notes Status

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: MG1655
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: slyA+
  • Genotype of Experimental Strain : slyA(del)::kan
  • Reference Condition:
OMP:0000028

decreased number of pili

Fig. 9-yeast agglutination assay

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: MG1655
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: slyA+
  • Genotype of Experimental Strain : slyA(del)::kan
  • Reference Condition:
OMP:0000268

decreased gene expression

Fig. 2-decreased expression of FimB-LacZ fusion



Notes

References

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