Multiple co‐regulatory elements and IHF are necessary for the control of fimB expression in response to sialic acid and N‐acetylglucosamine in Escherichia coli K‐12

Expression of the FimB recombinase, and hence the OFF‐to‐ON switching of type 1 fimbriation in Escherichia coli, is inhibited by sialic acid (Neu5Ac) and by GlcNAc. NanR (Neu5Ac‐responsive) and NagC (GlcNAc‐6P‐responsive) activate fimB expression by binding to operators (ONR and ONC1 respectively) located more than 600 bp upstream of the fimB promoter within the large (1.4 kb) nanC‐fimB intergenic region. Here it is demonstrated that NagC binding to a second site (ONC2), located 212 bp closer to fimB, also controls fimB expression, and that integration host factor (IHF), which binds midway between ONC1 and ONC2, facilitates NagC binding to its two operator sites. In contrast, IHF does not enhance the ability of NanR to activate fimB expression in the wild‐type background. Neither sequences up to 820 bp upstream of ONR, nor those 270 bp downstream of ONC2, are required for activation by NanR and NagC. However, placing the NanR, IHF and NagC binding sites closer to the fimB promoter enhances the ability of the regulators to activate fimB expression. These results support a refined model for how two potentially key indicators of host inflammation, Neu5Ac and GlcNAc, regulate type 1 fimbriation.

[1]  E. Vimr,et al.  Convergent Pathways for Utilization of the Amino Sugars N-Acetylglucosamine,N-Acetylmannosamine, and N-Acetylneuraminic Acid by Escherichia coli , 1999, Journal of bacteriology.

[2]  C. Brinton.  Non-Flagellar Appendages of Bacteria , 1959, Nature.

[3]  M. W. van der Woude,et al.  Formation of DNA Methylation Patterns: Nonmethylated GATC Sequences in gut and papOperons , 1998, Journal of bacteriology.

[4]  M. S. McClain,et al.  Inversion-independent phase variation of type 1 fimbriae in Escherichia coli , 1993, Journal of bacteriology.

[5]  E. Vimr,et al.  Derived structure of the putative sialic acid transporter from Escherichia coli predicts a novel sugar permease domain , 1995, Journal of bacteriology.

[6]  K. Swinger,et al.  IHF and HU: flexible architects of bent DNA. , 2004, Current opinion in structural biology.

[7]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[8]  Annie Kolb,et al.  Nag repressor-operator interactions: protein-DNA contacts cover more than two turns of the DNA helix. , 1995, Journal of Molecular Biology.

[9]  M. S. McClain,et al.  Lrp stimulates phase variation of type 1 fimbriation in Escherichia coli K-12 , 1993, Journal of bacteriology.

[10]  E. Vimr,et al.  Identification of an inducible catabolic system for sialic acids (nan) in Escherichia coli , 1985, Journal of bacteriology.

[11]  J. Pinkner,et al.  Type 1 pilus‐mediated bacterial invasion of bladder epithelial cells , 2000, The EMBO journal.

[12]  J. Steitz,et al.  Identification of a sex-factor-affinity site in E. coli as gamma delta. , 1981, Cold Spring Harbor symposia on quantitative biology.

[13]  J. Boudeau,et al.  Type 1 pili‐mediated adherence of Escherichia coli strain LF82 isolated from Crohn's disease is involved in bacterial invasion of intestinal epithelial cells , 2001, Molecular microbiology.

[14]  W. Shafer,et al.  Integration Host Factor is required for FarR repression of the farAB‐encoded efflux pump of Neisseria gonorrhoeae , 2006, Molecular microbiology.

[15]  L. Hang,et al.  Toll-Like Receptor 4 Expression and Cytokine Responses in the Human Urinary Tract Mucosa , 2004, Infection and Immunity.

[16]  H. Andrews-Polymenis,et al.  CsgA is a pathogen‐associated molecular pattern of Salmonella enterica serotype Typhimurium that is recognized by Toll‐like receptor 2 , 2005, Molecular microbiology.

[17]  S. Langermann,et al.  Prevention of mucosal Escherichia coli infection by FimH-adhesin-based systemic vaccination. , 1997, Science.

[18]  M. T. Beck,et al.  Osmolarity and pH Growth Conditions Regulate fim Gene Transcription and Type 1 Pilus Expression in Uropathogenic Escherichia coli , 2002, Infection and Immunity.

[19]  I. Blomfield,et al.  The leucine-responsive regulatory protein binds to the fim switch to control phase variation of type 1 fimbrial expression in Escherichia coli K-12 , 1994, Journal of bacteriology.

[20]  Phoebe A Rice,et al.  Integration host factor: putting a twist on protein-DNA recognition. , 2003, Journal of molecular biology.

[21]  C. Bloch,et al.  A key role for type 1 pili in enterobacterial communicability , 1992, Molecular microbiology.

[22]  M. Chapman,et al.  GlcNAc-6P Levels Modulate the Expression of Curli Fibers by Escherichia coli , 2006, Journal of bacteriology.

[23]  C. Higgins,et al.  Fimbrial phase variation in Escherichia coli: dependence on integration host factor and homologies with other site-specific recombinases , 1987, Journal of bacteriology.

[24]  Jeffrey H. Miller.  Experiments in molecular genetics , 1972 .

[25]  Peter Fischer,et al.  The Unresponsive Bystander: Are Bystanders More Responsive in Dangerous Emergencies? , 2006 .

[26]  M. W. van der Woude,et al.  Phase variation of Ag43 in Escherichia coli: Dam‐dependent methylation abrogates OxyR binding and OxyR‐mediated repression of transcription , 2000, Molecular microbiology.

[27]  I. Blomfield,et al.  Integration host factor stimulates both FimB‐ and FimE‐mediated site‐specific DNA inversion that controls phase variation of type 1 fimbriae expression in Escherichia coli , 1997, Molecular microbiology.

[28]  J. Plumbridge,et al.  Integrated regulatory responses of fimB to N-acetylneuraminic (sialic) acid and GlcNAc in Escherichia coli K-12. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[29]  M. Burdick,et al.  Hyaluronan (HA) fragments induce chemokine gene expression in alveolar macrophages. The role of HA size and CD44. , 1996, The Journal of clinical investigation.

[30]  V. Shingler,et al.  (p)ppGpp regulates type 1 fimbriation of Escherichia coli by modulating the expression of the site‐specific recombinase FimB , 2006, Molecular microbiology.

[31]  M. S. McClain,et al.  Roles of fimB and fimE in site-specific DNA inversion associated with phase variation of type 1 fimbriae in Escherichia coli , 1991, Journal of bacteriology.

[32]  M. Noordewier,et al.  Stress-induced DNA duplex destabilization (SIDD) in the E. coli genome: SIDD sites are closely associated with promoters. , 2004, Genome research.

[33]  E. Vimr,et al.  Diversity of Microbial Sialic Acid Metabolism , 2004, Microbiology and Molecular Biology Reviews.

[34]  J. Plumbridge,et al.  DNA binding sites for the Mlc and NagC proteins: regulation of nagE, encoding the N-acetylglucosamine-specific transporter in Escherichia coli. , 2001, Nucleic acids research.

[35]  Vincent Munster,et al.  Dam‐dependent phase variation of Ag43 in Escherichia coli is altered in a seqA mutant , 2002, Molecular microbiology.

[36]  Craig J. Benham,et al.  Activation of Gene Expression by a Novel DNA Structural Transmission Mechanism That Requires Supercoiling-induced DNA Duplex Destabilization in an Upstream Activating Sequence* , 1998, The Journal of Biological Chemistry.

[37]  I. Blomfield,et al.  Interaction of FimB and FimE with the fim switch that controls the phase variation of type 1 fimbriae in Escherichia coli K‐12 , 1996, Molecular microbiology.

[38]  K. Appelt,et al.  Purification of the Escherichia coli integration host factor (IHF) in one chromatographic step. , 1994, Gene.

[39]  E. Vimr,et al.  Regulation of Sialic Acid Catabolism by the DNA Binding Protein NanR in Escherichia coli , 2003, Journal of bacteriology.

[40]  S. Abraham,et al.  Mast cell modulation of neutrophil influx and bacterial clearance at sites of infection through TNF-α , 1996, Nature.

[41]  D. Friedman,et al.  Integration host factor is required for the DNA inversion that controls phase variation in Escherichia coli. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[42]  O. Pellegrini,et al.  Expression of the chitobiose operon of Escherichia coli is regulated by three transcription factors: NagC, ChbR and CAP , 2004, Molecular microbiology.

[43]  I. Blomfield,et al.  Allelic exchange in Escherichia coli using the Bacillus subtilis sacB gene and a temperature‐sensitive pSC101 replicon , 1991, Molecular microbiology.

[44]  I. Blomfield,et al.  Environmental regulation of the fim switch controlling type 1 fimbrial phase variation in Escherichia coli K-12: effects of temperature and media , 1993, Journal of bacteriology.

[45]  C. Svanborg,et al.  Type 1 fimbrial expression enhances Escherichia coli virulence for the urinary tract. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[46]  C. Tang,et al.  Type 1 fimbriae and extracellular polysaccharides are preeminent uropathogenic Escherichia coli virulence determinants in the murine urinary tract , 2002, Molecular microbiology.

[47]  W. McClure,et al.  Searching for and predicting the activity of sites for DNA binding proteins: compilation and analysis of the binding sites for Escherichia coli integration host factor (IHF). , 1990, Nucleic acids research.

[48]  J. Plumbridge,et al.  Function and Expression of an N-Acetylneuraminic Acid-Inducible Outer Membrane Channel in Escherichia coli , 2005, Journal of bacteriology.

[49]  Joe E Grissom,et al.  Carbon nutrition of Escherichia coli in the mouse intestine. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[50]  I. Blomfield,et al.  Leucine alters the interaction of the leucine‐responsive regulatory protein (Lrp) with the fim switch to stimulate site‐specific recombination in Escherichia coli , 1998, Molecular microbiology.

[51]  S. Hultgren,et al.  CD14- and Toll-Like Receptor-Dependent Activation of Bladder Epithelial Cells by Lipopolysaccharide and Type 1 Piliated Escherichia coli , 2003, Infection and Immunity.

[52]  H. Mobley,et al.  Transcriptome of Uropathogenic Escherichia coli during Urinary Tract Infection , 2004, Infection and Immunity.

[53]  S. Akira,et al.  Mechanism of pathogen‐specific TLR4 activation in the mucosa: Fimbriae, recognition receptors and adaptor protein selection , 2006, European journal of immunology.

[54]  S. Rigali,et al.  Subdivision of the Helix-Turn-Helix GntR Family of Bacterial Regulators in the FadR, HutC, MocR, and YtrA Subfamilies* , 2002, The Journal of Biological Chemistry.

[55]  G W Hatfield,et al.  Inhibition of DNA Supercoiling-dependent Transcriptional Activation by a Distant B-DNA to Z-DNA Transition* , 1999, The Journal of Biological Chemistry.

[56]  Phoebe A Rice,et al.  Crystal Structure of an IHF-DNA Complex: A Protein-Induced DNA U-Turn , 1996, Cell.

[57]  F. Neidhardt,et al.  Culture Medium for Enterobacteria , 1974, Journal of bacteriology.

[58]  S. El-Labany,et al.  Distant cis‐active sequences and sialic acid control the expression of fimB in Escherichia coli K‐12 , 2003, Molecular microbiology.

[59]  Xiangwu Nou,et al.  Regulation of pyelonephritis‐associated pili phase‐variation in Escherichia coli: binding of the Papl and the Lrp regulatory proteins is controlled by DNA methylation , 1993, Molecular microbiology.

[60]  B. Washburn,et al.  New method for generating deletions and gene replacements in Escherichia coli , 1989, Journal of bacteriology.

[61]  L. Enquist,et al.  Experiments With Gene Fusions , 1984 .

[62]  J. Abraham,et al.  An invertible element of DNA controls phase variation of type 1 fimbriae of Escherichia coli. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[63]  M. Horton,et al.  Hyaluronan Fragments Act as an Endogenous Danger Signal by Engaging TLR21 , 2006, The Journal of Immunology.

[64]  L. Hang,et al.  Type 1 fimbriae deliver an LPS‐ and TLR4‐dependent activation signal to CD14‐negative cells , 2001, Molecular microbiology.

[65]  R. Gumport,et al.  Genetic analysis of Escherichia coli integration host factor interactions with its bacteriophage lambda H' recognition site , 1991, Journal of bacteriology.

[66]  P. Klemm,et al.  Two regulatory fim genes, fimB and fimE, control the phase variation of type 1 fimbriae in Escherichia coli. , 1986, The EMBO journal.

[67]  A. Kolb,et al.  DNA bending and expression of the divergent nagE-B operons. , 1998, Nucleic acids research.

[68]  J. Plumbridge,et al.  Co‐ordinated regulation of amino sugar biosynthesis and degradation: the NagC repressor acts as both an activator and a repressor for the transcription of the glmUS operon and requires two separated NagC binding sites. , 1995, The EMBO journal.

[69]  C. Svanborg,et al.  Role of fimbriae‐mediated adherence for neutrophil migration across Escherichia coli‐infected epithelial cell layers , 1998, Molecular microbiology.

[70]  M. Lahooti,et al.  Modulation of the Sensitivity of FimB Recombination to Branched-Chain Amino Acids and Alanine in Escherichia coli K-12 , 2005, Journal of bacteriology.