Extensive Genomic Diversity in Pathogenic Escherichia coli and Shigella Strains Revealed by Comparative Genomic Hybridization Microarray

ABSTRACT Escherichia coli, including the closely related genus Shigella, is a highly diverse species in terms of genome structure. Comparative genomic hybridization (CGH) microarray analysis was used to compare the gene content of E. coli K-12 with the gene contents of pathogenic strains. Missing genes in a pathogen were detected on a microarray slide spotted with 4,071 open reading frames (ORFs) of W3110, a commonly used wild-type K-12 strain. For 22 strains subjected to the CGH microarray analyses 1,424 ORFs were found to be absent in at least one strain. The common backbone of the E. coli genome was estimated to contain about 2,800 ORFs. The mosaic distribution of absent regions indicated that the genomes of pathogenic strains were highly diversified becasue of insertions and deletions. Prophages, cell envelope genes, transporter genes, and regulator genes in the K-12 genome often were not present in pathogens. The gene contents of the strains tested were recognized as a matrix for a neighbor-joining analysis. The phylogenic tree obtained was consistent with the results of previous studies. However, unique relationships between enteroinvasive strains and Shigella, uropathogenic, and some enteropathogenic strains were suggested by the results of this study. The data demonstrated that the CGH microarray technique is useful not only for genomic comparisons but also for phylogenic analysis of E. coli at the strain level.

[1]  S M Payne,et al.  Complete Genome Sequence and Comparative Genomics of Shigella flexneri Serotype 2a Strain 2457T , 2003, Infection and Immunity.

[2]  C. Buchrieser,et al.  Analysis of Genome Plasticity in Pathogenic and Commensal Escherichia coli Isolates by Use of DNA Arrays , 2003, Journal of bacteriology.

[3]  G. Dougan,et al.  Genomic Comparison of Salmonella enterica Serovars and Salmonella bongori by Use of an S. enterica Serovar Typhimurium DNA Microarray , 2003, Journal of bacteriology.

[4]  Natalia Ivanova,et al.  The ERGOTM genome analysis and discovery system , 2003, Nucleic Acids Res..

[5]  F. Blattner,et al.  Extensive mosaic structure revealed by the complete genome sequence of uropathogenic Escherichia coli , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[6]  Yung-Sheng Chang,et al.  Regulation of the Hydrogenase-4 Operon of Escherichia coli by the σ54-Dependent Transcriptional Activators FhlA and HyfR , 2002, Journal of bacteriology.

[7]  Stanley Falkow,et al.  Improved analytical methods for microarray-based genome-composition analysis , 2002, Genome Biology.

[8]  Jie Dong,et al.  Genome sequence of Shigella flexneri 2a: insights into pathogenicity through comparison with genomes of Escherichia coli K12 and O157. , 2002, Nucleic acids research.

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

[10]  S. Porwollik,et al.  Evolutionary genomics of Salmonella: Gene acquisitions revealed by microarray analysis , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[11]  Nina Salama,et al.  Comparison of Genetic Divergence and Fitness between Two Subclones of Helicobacter pylori , 2001, Infection and Immunity.

[12]  S Falkow,et al.  Helicobacter pylori genetic diversity within the gastric niche of a single human host , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[13]  K. Kurokawa,et al.  Diversification of Escherichia coli genomes: are bacteriophages the major contributors? , 2001, Trends in microbiology.

[14]  T. Whittam,et al.  Pathogenesis and evolution of virulence in enteropathogenic and enterohemorrhagic Escherichia coli. , 2001, The Journal of clinical investigation.

[15]  M. Blaser,et al.  Helicobacter pylori strain-specific differences in genetic content, identified by microarray, influence host inflammatory responses. , 2001, The Journal of clinical investigation.

[16]  N. W. Davis,et al.  Genome sequence of enterohaemorrhagic Escherichia coli O157:H7 , 2001, Nature.

[17]  M. Hattori,et al.  Complete genome sequence of enterohemorrhagic Escherichia coli O157:H7 and genomic comparison with a laboratory strain K-12. , 2001, DNA research : an international journal for rapid publication of reports on genes and genomes.

[18]  G. Sherlock,et al.  A whole-genome microarray reveals genetic diversity among Helicobacter pylori strains. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[19]  H. Ochman,et al.  Evolutionary dynamics of full genome content in Escherichia coli , 2000, The EMBO journal.

[20]  G. Pupo,et al.  Multiple independent origins of Shigella clones of Escherichia coli and convergent evolution of many of their characteristics. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[21]  Alyssa C. Bumbaugh,et al.  Parallel evolution of virulence in pathogenic Escherichia coli , 2000, Nature.

[22]  Ash A. Alizadeh,et al.  Genome-wide analysis of DNA copy-number changes using cDNA microarrays , 1999, Nature Genetics.

[23]  D. Dykhuizen,et al.  Pathoadaptive mutations: gene loss and variation in bacterial pathogens. , 1999, Trends in microbiology.

[24]  R. W. Davis,et al.  A low rate of nucleotide changes in Escherichia coli K‐12 estimated from a comparison of the genome sequences between two different substrains , 1999, FEBS letters.

[25]  S. Teukolsky,et al.  Black Holes , 1998, gr-qc/9808035.

[26]  C. Bloch,et al.  "Black holes" and bacterial pathogenicity: a large genomic deletion that enhances the virulence of Shigella spp. and enteroinvasive Escherichia coli. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

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

[28]  G. Pupo,et al.  Evolutionary relationships among pathogenic and nonpathogenic Escherichia coli strains inferred from multilocus enzyme electrophoresis and mdh sequence studies , 1997, Infection and immunity.

[29]  P. Reeves,et al.  Organization of the Escherichia coli K-12 gene cluster responsible for production of the extracellular polysaccharide colanic acid , 1996, Journal of bacteriology.

[30]  J. Mattick,et al.  Escherichia coli contains a set of genes homologous to those involved in protein secretion, DNA uptake and the assembly of type-4 fimbriae in other bacteria. , 1994, Gene.

[31]  C. Sasakawa,et al.  The absence of a surface protease, OmpT, determines the intercellular spreading ability of Shigella: the relationship between the ompT and kcpA loci , 1993, Molecular microbiology.

[32]  B. Bochner Sleuthing out bacterial identities , 1989, Nature.

[33]  F. Neidhardt,et al.  Escherichia Coli and Salmonella: Typhimurium Cellular and Molecular Biology , 1987 .

[34]  S. T. Cowan Bergey's Manual of Determinative Bacteriology , 1948, Nature.

[35]  James T. Staley,et al.  Bergey's Manual of Determinative Bacteriology , 1939 .

[36]  S M Payne,et al.  Complete Genome Sequence and Comparative Genomics of Shigella flexneri Serotype 2a Strain 2457T , 2003, Infection and Immunity.

[37]  C. Buchrieser,et al.  Analysis of Genome Plasticity in Pathogenic and Commensal Escherichia coli Isolates by Use of DNA Arrays , 2003, Journal of bacteriology.

[38]  G. Dougan,et al.  Genomic Comparison of Salmonella enterica Serovars and Salmonella bongori by Use of an S. enterica Serovar Typhimurium DNA Microarray , 2003, Journal of bacteriology.

[39]  Natalia Ivanova,et al.  The ERGOTM genome analysis and discovery system , 2003, Nucleic Acids Res..

[40]  F. Blattner,et al.  Extensive mosaic structure revealed by the complete genome sequence of uropathogenic Escherichia coli , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[41]  Yung-Sheng Chang,et al.  Regulation of the Hydrogenase-4 Operon of Escherichia coli by the σ54-Dependent Transcriptional Activators FhlA and HyfR , 2002, Journal of bacteriology.

[42]  Stanley Falkow,et al.  Improved analytical methods for microarray-based genome-composition analysis , 2002, Genome Biology.

[43]  Jie Dong,et al.  Genome sequence of Shigella flexneri 2a: insights into pathogenicity through comparison with genomes of Escherichia coli K12 and O157. , 2002, Nucleic acids research.

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

[45]  S. Porwollik,et al.  Evolutionary genomics of Salmonella: Gene acquisitions revealed by microarray analysis , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[46]  Nina Salama,et al.  Comparison of Genetic Divergence and Fitness between Two Subclones of Helicobacter pylori , 2001, Infection and Immunity.

[47]  S Falkow,et al.  Helicobacter pylori genetic diversity within the gastric niche of a single human host , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[48]  K. Kurokawa,et al.  Diversification of Escherichia coli genomes: are bacteriophages the major contributors? , 2001, Trends in microbiology.

[49]  T. Whittam,et al.  Pathogenesis and evolution of virulence in enteropathogenic and enterohemorrhagic Escherichia coli. , 2001, The Journal of clinical investigation.

[50]  M. Blaser,et al.  Helicobacter pylori strain-specific differences in genetic content, identified by microarray, influence host inflammatory responses. , 2001, The Journal of clinical investigation.

[51]  M. Hattori,et al.  Complete genome sequence of enterohemorrhagic Escherichia coli O157:H7 and genomic comparison with a laboratory strain K-12. , 2001, DNA research : an international journal for rapid publication of reports on genes and genomes.

[52]  G. Sherlock,et al.  A whole-genome microarray reveals genetic diversity among Helicobacter pylori strains. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[53]  H. Ochman,et al.  Evolutionary dynamics of full genome content in Escherichia coli , 2000, The EMBO journal.

[54]  G. Pupo,et al.  Multiple independent origins of Shigella clones of Escherichia coli and convergent evolution of many of their characteristics. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[55]  Alyssa C. Bumbaugh,et al.  Parallel evolution of virulence in pathogenic Escherichia coli , 2000, Nature.

[56]  D. Dykhuizen,et al.  Pathoadaptive mutations: gene loss and variation in bacterial pathogens. , 1999, Trends in microbiology.

[57]  R. W. Davis,et al.  A low rate of nucleotide changes in Escherichia coli K‐12 estimated from a comparison of the genome sequences between two different substrains , 1999, FEBS letters.

[58]  C. Bloch,et al.  "Black holes" and bacterial pathogenicity: a large genomic deletion that enhances the virulence of Shigella spp. and enteroinvasive Escherichia coli. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

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

[60]  G. Pupo,et al.  Evolutionary relationships among pathogenic and nonpathogenic Escherichia coli strains inferred from multilocus enzyme electrophoresis and mdh sequence studies , 1997, Infection and immunity.

[61]  P. Reeves,et al.  Organization of the Escherichia coli K-12 gene cluster responsible for production of the extracellular polysaccharide colanic acid , 1996, Journal of bacteriology.

[62]  J. Mattick,et al.  Escherichia coli contains a set of genes homologous to those involved in protein secretion, DNA uptake and the assembly of type-4 fimbriae in other bacteria. , 1994, Gene.

[63]  C. Sasakawa,et al.  The absence of a surface protease, OmpT, determines the intercellular spreading ability of Shigella: the relationship between the ompT and kcpA loci , 1993, Molecular microbiology.

[64]  B. Bochner Sleuthing out bacterial identities , 1989, Nature.

[65]  F. Neidhardt,et al.  Escherichia Coli and Salmonella: Typhimurium Cellular and Molecular Biology , 1987 .

[66]  S. T. Cowan Bergey's Manual of Determinative Bacteriology , 1948, Nature.