Distant cis‐active sequences and sialic acid control the expression of fimB in Escherichia coli K‐12

The phase variation of type 1 fimbriation in Escherichia coli is controlled by the inversion of a 314 bp element of DNA, determined by FimB (switching in both directions) or FimE (switching from the ON‐to‐OFF orientation predominantly), and influenced by auxiliary factors IHF, Lrp and H‐NS. The fimB gene is separated from the divergently transcribed yjhATS operon by a large (1.4 kbp) intergenic region of unknown function. Here, we show that fimB expression is regulated by multiple cis‐active sequences that lie far upstream (>600 bp) of the transcription start sites for the recombinase gene. Two regions characterized further (regions 1 and 2) show sequence identity, and each coincides with a methylation‐protected Dam (5′‐GATC) site. Regions 1 and 2 apparently control fimB expression by an antirepression mechanism that involves additional sequences proximal to yjhA. Region 1 encompasses a 27 bp DNA sequence conserved upstream of genes known (nanAT ) or suspected (yjhBC) to be involved in sialic acid metabolism, and we show that FimB expression and recombination are suppressed by N‐acetylneuraminic acid. We propose that E. coli recognizes the amino sugars as a harbinger of potential host defence activation, and suppresses the expression of type 1 fimbriae in response.

[1]  A. Bäumler,et al.  Phase variation of the lpf operon is a mechanism to evade cross-immunity between Salmonella serotypes. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[2]  R. Goldman,et al.  Mannose-binding activity of Escherichia coli: a determinant of attachment and ingestion of the bacteria by macrophages. , 1980, Infection and immunity.

[3]  H. Seifert,et al.  Growth conditions mediate differential transcription of fim genes involved in phase variation of type 1 pili , 1992, Journal of bacteriology.

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

[5]  H. Seifert,et al.  Analysis of the fimB promoter region involved in type 1 pilus phase variation in Escherichia coli , 1994, Molecular and General Genetics MGG.

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

[7]  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.

[8]  F. Blattner,et al.  Analysis of the Escherichia coli genome. IV. DNA sequence of the region from 89.2 to 92.8 minutes. , 1993, Nucleic acids research.

[9]  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.

[10]  L. Maurer,et al.  A new locus, pilE, required for the binding of type 1 piliated Escherichia coli to erhythrocytes , 1985 .

[11]  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.

[12]  H. Mori,et al.  Genome‐wide analysis of deoxyadenosine methyltransferase‐mediated control of gene expression in Escherichia coli , 2002, Molecular microbiology.

[13]  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.

[14]  L. Blyn,et al.  Regulation of pap pilin phase variation by a mechanism involving differential dam methylation states. , 1990, The EMBO journal.

[15]  Hai-Young Wu,et al.  Long-range interaction between two promoters: Activation of the leu-500 promoter by a distant upstream promoter , 1995, Cell.

[16]  Lai-Xi Wang,et al.  Recruitment of Murine Neutrophils in Vivothrough Endogenous Sialidase Activity* , 2003, The Journal of Biological Chemistry.

[17]  Gina M. Donato,et al.  Promoter-specific repression of fimB expression by the Escherichia coli nucleoid-associated protein H-NS , 1997, Journal of bacteriology.

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

[19]  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.

[20]  M. S. McClain,et al.  Type 1 fimbriation and fimE mutants of Escherichia coli K-12 , 1991, Journal of bacteriology.

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

[22]  K. Dybvig,et al.  DNA rearrangements and phenotypic switching in prokaryotes , 1993, Molecular microbiology.

[23]  H. Mobley,et al.  In vivo phase variation of Escherichia coli type 1 fimbrial genes in women with urinary tract infection. , 1999, Infection and immunity.

[24]  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.

[25]  F. Blattner,et al.  Analysis of the Escherichia coli genome VI: DNA sequence of the region from 92.8 through 100 minutes. , 1995, Nucleic acids research.

[26]  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.

[27]  I. Jääskeläinen,et al.  Occurrence of sialic acids in healthy humans and different disorders , 1999, European journal of clinical investigation.

[28]  Y. Uchida,et al.  A new enzymatic method for the determination of sialic acid in serum and its application for a marker of acute phase reactants. , 1981, The Kobe journal of medical sciences.

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

[30]  P. Klemm,et al.  Localization of promoters in the fim gene cluster and the effect of H-NS on the transcription of fimB and fimE. , 1994, FEMS microbiology letters.

[31]  N. W. Davis,et al.  The complete genome sequence of Escherichia coli K-12. , 1997, Science.

[32]  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.

[33]  I. Blomfield,et al.  Orientational control of fimE expression in Escherichia coli , 2001, Molecular microbiology.

[34]  G. Church,et al.  A whole genome approach to in vivo DNA-protein interactions in E. coli , 1992, Nature.

[35]  A. N. Spiridonov,et al.  Congruent evolution of different classes of non-coding DNA in prokaryotic genomes. , 2002, Nucleic acids research.

[36]  I. Blomfield The regulation of pap and type 1 fimbriation in Escherichia coli. , 2001, Advances in microbial physiology.

[37]  D. Hartl,et al.  Diversifying Selection Governs Sequence Polymorphism in the Major Adhesin Proteins FimA, PapA, and SfaA of Escherichia coli , 1998, Journal of Molecular Evolution.

[38]  P. Valentin‐Hansen,et al.  Long-range cooperativity between gene regulatory sequences in a prokaryote , 1987, Nature.

[39]  B. Eisenstein Phase variation of type 1 fimbriae in Escherichia coli is under transcriptional control. , 1981, Science.

[40]  R. Simons,et al.  Improved single and multicopy lac-based cloning vectors for protein and operon fusions. , 1987, Gene.

[41]  Heidi J. Sofia,et al.  Analysis of the Escherichia coli genome. V. DNA sequence of the region from 76.0 to 81.5 minutes , 1993, Nucleic Acids Res..

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

[43]  I. Blomfield,et al.  The molecular basis for the specificity of fimE in the phase variation of type 1 fimbriae of Escherichia coli K‐12 , 1999, Molecular microbiology.

[44]  I. Blomfield,et al.  Bacterial Adhesion to Host Tissues: Regulation and function of phase variation in Escherichia coli , 2002 .

[45]  E. Vimr,et al.  To sialylate, or not to sialylate: that is the question. , 2002, Trends in microbiology.

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

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

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

[49]  D. Gally,et al.  Regulation of type 1 fimbrial expression in uropathogenic Escherichia coli : heterogeneity of expression through sequence changes in the fim switch region , 1998, Molecular microbiology.

[50]  A. Schaeffer,et al.  Regulation of production of type 1 pili among urinary tract isolates of Escherichia coli , 1986, Infection and Immunity.

[51]  M. Igo,et al.  A distant upstream site involved in the negative regulation of the Escherichia coli ompF gene , 1994, Journal of bacteriology.

[52]  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.

[53]  D. Low,et al.  Self-perpetuating epigenetic pili switches in bacteria , 2002, Proceedings of the National Academy of Sciences of the United States of America.

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

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

[56]  A. Ninfa,et al.  Initiation of transcription at the bacterial glnAp2 promoter by purified E. coli components is facilitated by enhancers , 1987, Cell.

[57]  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.