Distribution, Functional Expression, and Genetic Organization of Cif, a Phage-Encoded Type III-Secreted Effector from Enteropathogenic and Enterohemorrhagic Escherichia coli

ABSTRACT Enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli (EHEC) inject effector proteins into host cells via a type III secretion system encoded by the locus of enterocyte effacement (LEE). One of these effectors is Cif, encoded outside the LEE by a lambdoid prophage. In this study, we demonstrated that the Cif-encoding prophage of EPEC strain E22 is inducible and produces infectious phage particles. We investigated the distribution and functional expression of Cif in 5,049 E. coli strains of human, animal, and environmental origins. A total of 115 E. coli isolates from diverse origins and geographic locations carried cif. The presence of cif was tightly associated with the LEE, since all the cif-positive isolates were positive for the LEE. These results suggested that the Cif-encoding prophages have been widely disseminated within the natural population of E. coli but positively selected within the population of LEE-positive strains. Nonetheless, 66% of cif-positive E. coli strains did not induce a typical Cif-related phenotype in eukaryotic cells due to frameshift mutations or insertion of an IS element in the cif gene. The passenger region of the prophages carrying cif was highly variable and showed various combinations of IS elements and genes coding for other effectors such as nleB, nleC, nleH, nleG, espJ, and nleA/espI (some of which were also truncated). This diversity and the presence of nonfunctional effectors should be taken into account to assess EPEC and EHEC pathogenicity and tropism.

[1]  F. Taieb,et al.  Escherichia coli cyclomodulin Cif induces G2 arrest of the host cell cycle without activation of the DNA‐damage checkpoint‐signalling pathway , 2006, Cellular microbiology.

[2]  Tetsuya Hayashi,et al.  An extensive repertoire of type III secretion effectors in Escherichia coli O157 and the role of lambdoid phages in their dissemination , 2006, Proceedings of the National Academy of Sciences.

[3]  David S Guttman,et al.  Terminal Reassortment Drives the Quantum Evolution of Type III Effectors in Bacterial Pathogens , 2006, PLoS pathogens.

[4]  L. Beutin,et al.  Characterization of Shiga Toxin Gene (stx)-Positive and Intimin Gene (eae)-Positive Escherichia coli Isolates from Wastewater of Slaughterhouses in France , 2006, Applied and Environmental Microbiology.

[5]  A. Caprioli,et al.  Detection of toxB, a Plasmid Virulence Gene of Escherichia coli O157, in Enterohemorrhagic and Enteropathogenic E. coli , 2005, Journal of Clinical Microbiology.

[6]  G. Frankel,et al.  Enteropathogenic and Enterohemorrhagic Escherichia coli Infections: Translocation, Translocation, Translocation , 2005, Infection and Immunity.

[7]  Songnian Hu,et al.  The genome sequence of Salmonella enterica serovar Choleraesuis, a highly invasive and resistant zoonotic pathogen , 2005, Nucleic acids research.

[8]  F. Taieb,et al.  Cyclomodulins: bacterial effectors that modulate the eukaryotic cell cycle. , 2005, Trends in microbiology.

[9]  Huanming Yang,et al.  Pseudogene accumulation might promote the adaptive microevolution of Yersinia pestis. , 2005, Journal of medical microbiology.

[10]  W. Hardt,et al.  Bacteriophage-encoded type III effectors in Salmonella enterica subspecies 1 serovar Typhimurium. , 2005, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[11]  Herbert Schmidt,et al.  Shiga toxin-encoding bacteriophages--genomes in motion. , 2004, International journal of medical microbiology : IJMM.

[12]  A. Blanch,et al.  Diversity of stx2 converting bacteriophages induced from Shiga-toxin-producing Escherichia coli strains isolated from cattle. , 2004, Microbiology.

[13]  Wolf-Dietrich Hardt,et al.  Phages and the Evolution of Bacterial Pathogens: from Genomic Rearrangements to Lysogenic Conversion , 2004, Microbiology and Molecular Biology Reviews.

[14]  M. Gerstein,et al.  Comprehensive analysis of pseudogenes in prokaryotes: widespread gene decay and failure of putative horizontally transferred genes , 2004, Genome Biology.

[15]  É. Oswald,et al.  Identification of the Secretion and Translocation Domain of the Enteropathogenic and Enterohemorrhagic Escherichia coli Effector Cif, Using TEM-1 β-Lactamase as a New Fluorescence-Based Reporter , 2004, Journal of bacteriology.

[16]  Hung Bui,et al.  A specific genetic background is required for acquisition and expression of virulence factors in Escherichia coli. , 2004, Molecular biology and evolution.

[17]  Harry L. T. Mobley,et al.  Pathogenic Escherichia coli , 2004, Nature Reviews Microbiology.

[18]  J. Jofre,et al.  Abundance in sewage of bacteriophages infecting Escherichia coli O157:H7. , 2004, Methods in molecular biology.

[19]  X. Tu,et al.  Enteropathogenic and enterohaemorrhagic Escherichia coli deliver a novel effector called Cif, which blocks cell cycle G2/M transition , 2003, Molecular microbiology.

[20]  L. Beutin,et al.  Production of Cytolethal Distending Toxins by Pathogenic Escherichia coli Strains Isolated from Human and Animal Sources: Establishment of the Existence of a New cdt Variant (Type IV) , 2003, Journal of Clinical Microbiology.

[21]  A. Caprioli,et al.  A Mosaic Pathogenicity Island Made Up of the Locus of Enterocyte Effacement and a Pathogenicity Island of Escherichiacoli O157:H7 Is Frequently Present in Attaching and Effacing E. coli , 2003, Infection and Immunity.

[22]  Ken Kurokawa,et al.  Genomic diversity of enterohemorrhagic Escherichia coli O157 revealed by whole genome PCR scanning , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[23]  L. Beutin,et al.  Genetic Diversity of Intimin Genes of Attaching and Effacing Escherichia coli Strains , 2002, Journal of Clinical Microbiology.

[24]  Matthew K. Waldor,et al.  Bacteriophage Control of Bacterial Virulence , 2002, Infection and Immunity.

[25]  H. Schmidt,et al.  The Nucleotide Sequence of Shiga Toxin (Stx) 2e-Encoding Phage φP27 Is Not Related to Other Stx Phage Genomes, but the Modular Genetic Structure Is Conserved , 2002, Infection and Immunity.

[26]  A. Caprioli,et al.  Detection and characterization of class 1 integrons in enterohemorrhagic Escherichia coli. , 2002, Microbial drug resistance.

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

[28]  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.

[29]  O. Clermont,et al.  Rapid and Simple Determination of theEscherichia coli Phylogenetic Group , 2000, Applied and Environmental Microbiology.

[30]  A. Caprioli,et al.  Typing of Intimin Genes in Human and Animal Enterohemorrhagic and Enteropathogenic Escherichia coli: Characterization of a New Intimin Variant , 2000, Infection and Immunity.

[31]  A. Caprioli,et al.  Molecular characterisation of verocytotoxin-producing Escherichia coli of serogroup O111 from different countries. , 1999, Journal of medical microbiology.

[32]  H. Karch,et al.  Transduction of Enteric Escherichia coli Isolates with a Derivative of Shiga Toxin 2-Encoding Bacteriophage φ3538 Isolated from Escherichia coli O157:H7 , 1999, Applied and Environmental Microbiology.

[33]  A. Campbell,et al.  Integration specificities of two lambdoid phages (21 and e14) that insert at the same attB site , 1997, Journal of Bacteriology.

[34]  L. Bossi,et al.  Unsuspected prophage‐like elements in Salmonella typhimurium , 1997, Molecular microbiology.

[35]  B. China,et al.  Typing of bovine attaching and effacing Escherichia coli by multiplex in vitro amplification of virulence-associated genes , 1996, Applied and environmental microbiology.

[36]  T. Whittam,et al.  Clonal relationships among Escherichia coli strains that cause hemorrhagic colitis and infantile diarrhea , 1993, Infection and immunity.

[37]  G. Schoolnik,et al.  An inducible bundle-forming pilus of enteropathogenic Escherichia coli. , 1991, Science.

[38]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.

[39]  B. Bachmann,et al.  Derivations and genotypes of some mutant derivatives of Escherichia coli K12 , 1987 .

[40]  M. Thomas,et al.  Secondary attachment site for bacteriophage lambda in the guaB gene of Escherichia coli , 1986, Journal of bacteriology.

[41]  H. Ochman,et al.  Standard reference strains of Escherichia coli from natural populations , 1984, Journal of bacteriology.

[42]  R. Hoess,et al.  Structure of the lambda att sites generated by int-dependent deletions. , 1978, Proceedings of the National Academy of Sciences of the United States of America.