Genomic Analysis of a Pathogenicity Island in Uropathogenic Escherichia coli CFT073: Distribution of Homologous Sequences among Isolates from Patients with Pyelonephritis, Cystitis, and CatheterAssociated Bacteriuria and from Fecal Samples

ABSTRACT Urinary tract infection is the most frequently diagnosed kidney and urologic disease and Escherichia coli is by far the most common etiologic agent. Uropathogenic strains have been shown to contain blocks of DNA termed pathogenicity islands (PAIs) which contribute to their virulence. We have defined one of these regions of DNA within the chromosome of a highly virulentE. coli strain, CFT073, isolated from the blood and urine of a woman with acute pyelonephritis. The 57,988-bp stretch of DNA has characteristics which define PAIs, including a size greater than 30 kb, the presence of insertion sequences, distinct segmentation of K-12 and J96 origin, GC content (42.9%) different from that of total genomic DNA (50.8%), and the presence of virulence genes (hly and pap). Within this region, we have identified 44 open reading frames; of these 44, 10 are homologous to entries in the complete K-12 genome sequence, 4 are nearly identical to the sequences of E. coli J96 encoding the HlyA hemolysin, 11 encode P fimbriae, and 19 show no homology to J96 or K-12 entries. To determine whether sequences found within the junctions of the PAI of CFT073 were common to other uropathogenic strains ofE. coli, 11 probes were isolated along the length of the PAI and were hybridized to dot blots of genomic DNA isolated from clinical isolates (67 from patients with acute pyelonephritis, 38 from patients with cystitis, 49 from patients with catheter-associated bacteriuria, and 27 from fecal samples). These sequences were found significantly more often in strains associated with the clinical syndromes of acute pyelonephritis (79%) and cystitis (82%) than in those associated with catheter-associated bacteriuria (58%) and in fecal strains (22%) (P < 0.001). From these regions, we have identified a putative iron transport system and genes other than hly and pap that may contribute to the virulent phenotype of uropathogenic E. coli strains.

[1]  R. Deonier Native Insertion Sequence Elements : Locations , Distributions , and Sequence Relationships , 1999 .

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

[3]  H. Mobley,et al.  Pathogenicity island sequences of pyelonephritogenic Escherichia coli CFT073 are associated with virulent uropathogenic strains , 1997, Infection and immunity.

[4]  J. Mecsas,et al.  Molecular mechanisms of bacterial virulence: type III secretion and pathogenicity islands. , 1996, Emerging infectious diseases.

[5]  D. Berg,et al.  Two pathogenicity islands in uropathogenic Escherichia coli J96: cosmid cloning and sample sequencing , 1996, Infection and immunity.

[6]  Leeds Ca Pathogenicity islands and the evolution of bacterial pathogens. , 1996 .

[7]  C. Lee Pathogenicity islands and the evolution of bacterial pathogens. , 1996, Infectious agents and disease.

[8]  H. Mobley,et al.  Urinary tract infections : molecular pathogenesis and clinical management , 1996 .

[9]  K. Shanmugam,et al.  Genetic analysis of the modABCD (molybdate transport) operon of Escherichia coli , 1995, Journal of bacteriology.

[10]  W. Rabsch,et al.  tRNA genes and pathogenicity islands: influence on virulence and metabolic properties of uropathogenic Escherichia coli , 1995, Molecular microbiology.

[11]  B. Foxman,et al.  Bacterial virulence characteristics of Escherichia coli isolates from first-time urinary tract infection. , 1995, The Journal of infectious diseases.

[12]  A. Caprioli,et al.  Gene clusters encoding the cytotoxic necrotizing factor type 1, Prs-fimbriae and alpha-hemolysin form the pathogenicity island II of the uropathogenic Escherichia coli strain J96. , 1995, FEMS microbiology letters.

[13]  W. Jacobs,et al.  Identification of genes involved in the sequestration of iron in mycobacteria: the ferric exochelin biosynthetic and uptake pathways , 1994, Molecular microbiology.

[14]  K. Jarvis,et al.  Isogenic P‐fimbrial deletion mutants of pyelonephritogenic Escherichia coli: the role of α Gal(1–4)β Gal binding in virulence of a wild‐type strain , 1993 .

[15]  A Danchin,et al.  Bacillus subtilis genome project: cloning and sequencing of the 97 kb region from 325° to 333deg; , 1993, Molecular microbiology.

[16]  E. Ohtsubo,et al.  Distribution of the Shigella sonnei insertion elements in Enterobacteriaceae. , 1993, Gene.

[17]  M. Arnaud,et al.  Bacillus subtilis genome project: cloning and sequencing of the 97 kb region from 325 degrees to 333 degrees. , 1993, Molecular microbiology.

[18]  J. R. Johnson,et al.  Virulence factors in Escherichia coli urinary tract infection , 1991, Clinical Microbiology Reviews.

[19]  R. D'ari,et al.  A relationship between L-serine degradation and methionine biosynthesis in Escherichia coli K12. , 1990, Journal of general microbiology.

[20]  H. Mobley,et al.  Pyelonephritogenic Escherichia coli and killing of cultured human renal proximal tubular epithelial cells: role of hemolysin in some strains , 1990, Infection and immunity.

[21]  E. Ohtsubo,et al.  Complete sequence of IS629. , 1990, Nucleic acids research.

[22]  J Hacker,et al.  Genetic determinants coding for fimbriae and adhesins of extraintestinal Escherichia coli. , 1990, Current topics in microbiology and immunology.

[23]  B. Rak,et al.  Regulation of the bgl operon of Escherichia coli by transcriptional antitermination. , 1988, The EMBO journal.

[24]  E. Stokland,et al.  Virulence of Escherichia coli in relation to host factors in women with symptomatic urinary tract infection , 1988, Journal of clinical microbiology.

[25]  J. R. Johnson,et al.  Aerobactin and other virulence factor genes among strains of Escherichia coli causing urosepsis: association with patient characteristics , 1988, Infection and immunity.

[26]  M. B. Kaack,et al.  P-Fimbriated Escherichia coli Urinary Tract Infection: A Clinical Correlation , 1987, Southern medical journal.

[27]  S. Normark,et al.  Nucleotide sequence, regulation and functional analysis of the papC gene required for cell surface localization of Pap pili of uropathogenic Escherichia coli , 1987, Molecular microbiology.

[28]  E. Ohtsubo,et al.  Isolation and characterization of IS elements repeated in the bacterial chromosome. , 1987, Journal of molecular biology.

[29]  S. Normark,et al.  The PapG protein is the alpha-D-galactopyranosyl-(1----4)-beta-D-galactopyranose-binding adhesin of uropathogenic Escherichia coli. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[30]  S. Normark,et al.  Localization of the receptor-binding protein adhesin at the tip of the bacterial pilus , 1987, Nature.

[31]  B. Rak,et al.  Beta-glucoside (bgl) operon of Escherichia coli K-12: nucleotide sequence, genetic organization, and possible evolutionary relationship to regulatory components of two Bacillus subtilis genes , 1987, Journal of bacteriology.

[32]  M. Rhen,et al.  Comparison of the nucleotide sequences of the genes encoding the KS71A and F7(1) fimbrial antigens of uropathogenic Escherichia coli. , 1985, European journal of biochemistry.

[33]  S Falkow,et al.  Gal-Gal binding and hemolysin phenotypes and genotypes associated with uropathogenic Escherichia coli. , 1985, The New England journal of medicine.

[34]  R. Welch,et al.  Nucleotide sequence of an Escherichia coli chromosomal hemolysin , 1985, Journal of bacteriology.

[35]  H. Bergmans,et al.  Nucleotide sequence of the gene encoding the F72 fimbrial subunit of a uropathogenic Escherichia coli strain. , 1984, Gene.

[36]  W. Stamm,et al.  Role of fimbriated Escherichia coli in urinary tract infections in adult women: correlation with localization studies. , 1984, The Journal of infectious diseases.

[37]  G. Schoolnik,et al.  Nucleotide sequence of the papA gene encoding the Pap pilus subunit of human uropathogenic Escherichia coli , 1984, Journal of bacteriology.

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

[39]  S Falkow,et al.  Genetics of digalactoside-binding adhesin from a uropathogenic Escherichia coli strain , 1983, Infection and immunity.

[40]  R. Freter,et al.  Ascending, unobstructed urinary tract infection in mice caused by pyelonephritogenic Escherichia coli of human origin , 1983, Infection and immunity.

[41]  W. Anthony,et al.  A prospective microbiologic study of bacteriuria in patients with chronic indwelling urethral catheters. , 1982, The Journal of infectious diseases.

[42]  F C Kafatos,et al.  Determination of nucleic acid sequence homologies and relative concentrations by a dot hybridization procedure. , 1979, Nucleic acids research.

[43]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.