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Article: Enteropathogenic and enterohemorrhagic Escherichia coli infections: translocation, translocation, translocation.

Garmendia, Junkal / Frankel, Gad / Crepin, Valérie F

2005 Volume 73, Issue 5, Page(s) 2573–2585

MeSH term(s)	Animals ; Diarrhea/microbiology ; Diarrhea/physiopathology ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Escherichia coli/pathogenicity ; Escherichia coli Infections/microbiology ; Escherichia coli Infections/physiopathology ; Escherichia coli O157/genetics ; Escherichia coli O157/metabolism ; Escherichia coli O157/pathogenicity ; Escherichia coli Proteins/genetics ; Escherichia coli Proteins/metabolism ; Humans ; Intestines/microbiology ; Intestines/pathology ; Mice ; Protein Transport
Chemical Substances	Escherichia coli Proteins
Language	English
Publishing date	2005-05
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	218698-6
ISSN	1098-5522 ; 0019-9567
ISSN (online)	1098-5522
ISSN	0019-9567
DOI	10.1128/IAI.73.5.2573-2585.2005
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Article: Enteropathogenic and Enterohemorrhagic Escherichia coli Infections: Translocation, Translocation, Translocation

Garmendia, Junkal / Frankel, Gad / Crepin, Valérie F

Infection and immunity. 2005 May., v. 73, no. 5

2005

Language	English
Dates of publication	2005-05
Size	p. 2573-2585.
Document type	Article
ZDB-ID	218698-6
ISSN	1098-5522 ; 0019-9567
ISSN (online)	1098-5522
ISSN	0019-9567
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Article ; Online: Identification of Translocation Inhibitors Targeting the Type III Secretion System of Enteropathogenic Escherichia coli.

Mühlen, Sabrina / Zapol'skii, Viktor A / Bilitewski, Ursula / Dersch, Petra

Antimicrobial agents and chemotherapy

2021 Volume 65, Issue 12, Page(s) e0095821

Abstract: Infections with enteropathogenic Escherichia coli (EPEC) cause severe diarrhea in children ... enterohemorrhagic E. coli infections. ... libraries and identified three substances which showed a significant dose-dependent effect on translocation ...

Abstract	Infections with enteropathogenic Escherichia coli (EPEC) cause severe diarrhea in children. The noninvasive bacteria adhere to enterocytes of the small intestine and use a type III secretion system (T3SS) to inject effector proteins into host cells to modify and exploit cellular processes in favor of bacterial survival and replication. Several studies have shown that the T3SSs of bacterial pathogens are essential for virulence. Furthermore, the loss of T3SS-mediated effector translocation results in increased immune recognition and clearance of the bacteria. The T3SS is, therefore, considered a promising target for antivirulence strategies and novel therapeutics development. Here, we report the results of a high-throughput screening assay based on the translocation of the EPEC effector protein Tir (translocated intimin receptor). Using this assay, we screened more than 13,000 small molecular compounds of six different compound libraries and identified three substances which showed a significant dose-dependent effect on translocation without adverse effects on bacterial or eukaryotic cell viability. In addition, these substances reduced bacterial binding to host cells, effector-dependent cell detachment, and abolished attaching and effacing lesion formation without affecting the expression of components of the T3SS or associated effector proteins. Moreover, no effects of the inhibitors on bacterial motility or Shiga-toxin expression were observed. In summary, we have identified three new compounds that strongly inhibit T3SS-mediated translocation of effectors into mammalian cells, which could be valuable as lead substances for treating EPEC and enterohemorrhagic E. coli infections.
MeSH term(s)	Animals ; Enteropathogenic Escherichia coli ; Escherichia coli Infections/drug therapy ; Escherichia coli Proteins/genetics ; HeLa Cells ; Humans ; Type III Secretion Systems/genetics ; Virulence
Chemical Substances	Escherichia coli Proteins ; Type III Secretion Systems
Language	English
Publishing date	2021-09-20
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	217602-6
ISSN	1098-6596 ; 0066-4804
ISSN (online)	1098-6596
ISSN	0066-4804
DOI	10.1128/AAC.00958-21
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Article ; Online: Effects of Psidium guajava leaf extract on secretion systems of Gram-negative enteropathogenic bacteria.

Nakasone, Noboru / Ogura, Yasunori / Higa, Naomi / Toma, Claudia / Koizumi, Yukiko / Takaesu, Giichi / Suzuki, Toshihiko / Yamashiro, Tetsu

Microbiology and immunology

2018

Abstract: ... the function of the type III secretion system (T3SS) of enteropathogenic Escherichia coli (EPEC) and ... treatment and prevention of gram-negative enteropathogenic bacterial infections. ... enterohemorrhagic E. coli (EHEC). Among candidates examined, we found that an extract from the leaves ...

Abstract	We screened a total of 672 plant-tissue extracts to search for phytochemicals that inhibit the function of the type III secretion system (T3SS) of enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli (EHEC). Among candidates examined, we found that an extract from the leaves of Psidium guajava (guava) inhibited the secretion of the EspB protein from EPEC and EHEC without affecting bacterial growth. The guava extract (GE) also inhibited EPEC and EHEC from adhering to and injecting EspB protein into HEp-2 cells. GE seemed to block the translocation of EspB from the bacterial cells to the culture medium. In addition to EPEC and EHEC, GE also inhibited the T3SS of Yersinia pseudotuberculosis and Salmonella enterica serovar Typhimurium. After exposure to GE, Y. pseudotuberculosis stopped the secretion of Yop proteins and lost its ability to induce the apoptosis of mouse bone marrow-derived macrophages. S. Typhimurium exposed to GE ceased the secretion of Sip proteins and lost its ability to invade HEp-2 cells. GE inhibited EspC secretion, the type V secretion protein of EPEC, but not Shiga toxin2 from EHEC. Thus, our results suggest that guava leaves contain a novel type of antimicrobial compound that could be used for the therapeutic treatment and prevention of gram-negative enteropathogenic bacterial infections.
Language	English
Publishing date	2018-05-23
Publishing country	Australia
Document type	Journal Article
ZDB-ID	224792-6
ISSN	1348-0421 ; 0385-5600
ISSN (online)	1348-0421
ISSN	0385-5600
DOI	10.1111/1348-0421.12604
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Article ; Online: Dynamics of the type III secretion system activity of enteropathogenic Escherichia coli.

Mills, Erez / Baruch, Kobi / Aviv, Gili / Nitzan, Mor / Rosenshine, Ilan

mBio

2013 Volume 4, Issue 4

Abstract: ... enterohemorrhagic Escherichia coli (EHEC) represent a global public health problem. New strategies to combat EPEC and EHEC infections ... based translocation assay, focusing mainly on enteropathogenic Escherichia coli (EPEC) and partly ... of the translocating bacteria, translocation temporal order, and relative amounts of each of the translocated effectors ...

Abstract	Unlabelled: Type III secretion systems (TTSSs) are employed by pathogens to translocate host cells with effector proteins, which are crucial for virulence. The dynamics of effector translocation, behavior of the translocating bacteria, translocation temporal order, and relative amounts of each of the translocated effectors are all poorly characterized. To address these issues, we developed a microscopy-based assay that tracks effector translocation. We used this assay alongside a previously described real-time population-based translocation assay, focusing mainly on enteropathogenic Escherichia coli (EPEC) and partly comparing it to Salmonella. We found that the two pathogens exhibit different translocation behaviors: in EPEC, a subpopulation that formed microcolonies carried out most of the translocation activity, while Salmonella executed protein translocation as planktonic bacteria. We also noted variability in host cell susceptibility, with some cells highly resistant to translocation. We next extended the study to determine the translocation dynamics of twenty EPEC effectors and found that all exhibited distinct levels of translocation efficiency. Further, we mapped the global effects of key TTSS-related components on TTSS activity. Our results provide a comprehensive description of the dynamics of the TTSS activity of EPEC and new insights into the mechanisms that control the dynamics. Importance: EPEC and the closely related enterohemorrhagic Escherichia coli (EHEC) represent a global public health problem. New strategies to combat EPEC and EHEC infections are needed, and development of such strategies requires better understanding of their virulence machinery. The TTSS is a critical virulence mechanism employed by these pathogens, and by others, including Salmonella. In this study, we aimed at elucidating new aspects of TTSS function. The results obtained provide a comprehensive description of the dynamics of TTSS activity of EPEC and new insights into the mechanisms that control these changes. This knowledge sets the stage for further analysis of the system and may accelerate the development of new ways to treat EPEC and EHEC infections. Further, the newly described microscopy-based assay can be readily adapted to study the dynamics of TTSS activity in other pathogens.
MeSH term(s)	Bacterial Proteins/metabolism ; Bacterial Secretion Systems ; Enteropathogenic Escherichia coli/metabolism ; Epithelial Cells/microbiology ; HeLa Cells ; Humans ; Protein Transport ; Salmonella/metabolism ; Virulence Factors/metabolism
Chemical Substances	Bacterial Proteins ; Bacterial Secretion Systems ; Virulence Factors
Language	English
Publishing date	2013-07-30
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2557172-2
ISSN	2150-7511 ; 2161-2129
ISSN (online)	2150-7511
ISSN	2161-2129
DOI	10.1128/mBio.00303-13
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Article: Dynamics of the Type III Secretion System Activity of Enteropathogenic Escherichia coli

Mills, Erez / Baruch, Kobi / Aviv, Gili / Nitzan, Mor / Rosenshine, Ilan

mBio. 2013 Aug. 30, v. 4, no. 4

2013

Abstract	Type III secretion systems (TTSSs) are employed by pathogens to translocate host cells with effector proteins, which are crucial for virulence. The dynamics of effector translocation, behavior of the translocating bacteria, translocation temporal order, and relative amounts of each of the translocated effectors are all poorly characterized. To address these issues, we developed a microscopy-based assay that tracks effector translocation. We used this assay alongside a previously described real-time population-based translocation assay, focusing mainly on enteropathogenic Escherichia coli (EPEC) and partly comparing it to Salmonella . We found that the two pathogens exhibit different translocation behaviors: in EPEC, a subpopulation that formed microcolonies carried out most of the translocation activity, while Salmonella executed protein translocation as planktonic bacteria. We also noted variability in host cell susceptibility, with some cells highly resistant to translocation. We next extended the study to determine the translocation dynamics of twenty EPEC effectors and found that all exhibited distinct levels of translocation efficiency. Further, we mapped the global effects of key TTSS-related components on TTSS activity. Our results provide a comprehensive description of the dynamics of the TTSS activity of EPEC and new insights into the mechanisms that control the dynamics. IMPORTANCE EPEC and the closely related enterohemorrhagic Escherichia coli (EHEC) represent a global public health problem. New strategies to combat EPEC and EHEC infections are needed, and development of such strategies requires better understanding of their virulence machinery. The TTSS is a critical virulence mechanism employed by these pathogens, and by others, including Salmonella . In this study, we aimed at elucidating new aspects of TTSS function. The results obtained provide a comprehensive description of the dynamics of TTSS activity of EPEC and new insights into the mechanisms that control these changes. This knowledge sets the stage for further analysis of the system and may accelerate the development of new ways to treat EPEC and EHEC infections. Further, the newly described microscopy-based assay can be readily adapted to study the dynamics of TTSS activity in other pathogens.
Keywords	Salmonella ; bacteria ; enterohemorrhagic Escherichia coli ; enteropathogenic Escherichia coli ; pathogens ; protein transport ; public health ; type III secretion system ; virulence
Language	English
Dates of publication	2013-0830
Size	p. e00303-13.
Publishing place	American Society for Microbiology
Document type	Article
ZDB-ID	2557172-2
ISSN	2150-7511 ; 2161-2129
ISSN (online)	2150-7511
ISSN	2161-2129
DOI	10.1128/mBio.00303-13
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Article ; Online: Enterohaemorrhagic, but not enteropathogenic, Escherichia coli infection of epithelial cells disrupts signalling responses to tumour necrosis factor-alpha.

Gareau, Mélanie G / Ho, Nathan K / Brenner, Dirk / Sousa, Andrew J / LeBourhis, Lionel / Mak, Tak W / Girardin, Stephen E / Philpott, Dana J / Sherman, Philip M

Microbiology (Reading, England)

2011 Volume 157, Issue Pt 10, Page(s) 2963–2973

Abstract: Enterohaemorrhagic Escherichia coli (EHEC), serotype O157 : H7 is a non-invasive ... of IκBα), immunofluorescence (p65 nuclear translocation) and immunoassay (CXCL-8 secretion), and each was ... following exposure to either enteropathogenic E. coli, strain E2348/69 (O127 : H6) or the laboratory E. coli ...

Abstract	Enterohaemorrhagic Escherichia coli (EHEC), serotype O157 : H7 is a non-invasive, pathogenic bacterium that employs a type III secretion system (T3SS) to inject effector proteins into infected cells. In this study, we demonstrate that EHEC blocks tumour necrosis factor-alpha (TNFα)-induced NF-κB signalling in infected epithelial cells. HEK293T and INT407 epithelial cells were challenged with EHEC prior to stimulation with TNFα. Using complementary techniques, stimulation with TNFα caused activation of NF-κB, as determined by luciferase reporter assay (increase in gene expression), Western blotting (phosphorylation of IκBα), immunofluorescence (p65 nuclear translocation) and immunoassay (CXCL-8 secretion), and each was blocked by EHEC O157 : H7 infection. In contrast, subversion of host cell signalling was not observed following exposure to either enteropathogenic E. coli, strain E2348/69 (O127 : H6) or the laboratory E. coli strain HB101. Heat-killed EHEC had no effect on NF-κB activation by TNFα. Inhibition was mediated, at least in part, by Shiga toxins and by the O157 plasmid, but not by the T3SS or flagellin, as demonstrated by using isogenic mutant strains. These findings indicate the potential for developing novel therapeutic targets to interrupt the infectious process.
MeSH term(s)	Cell Line/microbiology ; Enterohemorrhagic Escherichia coli/genetics ; Enterohemorrhagic Escherichia coli/physiology ; Epithelial Cells/metabolism ; Epithelial Cells/microbiology ; Escherichia coli Infections/metabolism ; Escherichia coli Infections/microbiology ; Escherichia coli O157/genetics ; Escherichia coli O157/physiology ; Humans ; NF-kappa B/metabolism ; Signal Transduction ; Tumor Necrosis Factor-alpha/metabolism
Chemical Substances	NF-kappa B ; Tumor Necrosis Factor-alpha
Language	English
Publishing date	2011-07-28
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1180712-x
ISSN	1465-2080 ; 1350-0872
ISSN (online)	1465-2080
ISSN	1350-0872
DOI	10.1099/mic.0.051094-0
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Article ; Online: Enteropathogenic and enterohemorrhagic Escherichia coli type III secretion effector EspV induces radical morphological changes in eukaryotic cells.

Arbeloa, Ana / Oates, Clare V / Marchès, Oliver / Hartland, Elizabeth L / Frankel, Gad

Infection and immunity

2010 Volume 79, Issue 3, Page(s) 1067–1076

Abstract: Enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic Escherichia coli (EHEC) are ... for subversion of signal transduction pathways and colonization of the mammalian gut mucosa. While a core set ... of effectors is conserved between EPEC and EHEC strains, a growing number of accessory effectors that were ...

Abstract	Enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic Escherichia coli (EHEC) are important human pathogens that rely on translocation of type III secretion system (T3SS) effectors for subversion of signal transduction pathways and colonization of the mammalian gut mucosa. While a core set of effectors is conserved between EPEC and EHEC strains, a growing number of accessory effectors that were found at various frequencies in clinical and environmental isolates have been recently identified. Recent genome projects identified espV as a pseudogene in EHEC but a putative functional gene in EPEC strains E110019 and E22 and the closely related mouse pathogen Citrobacter rodentium. The aim of this study was to determine the distribution of espV among clinical EPEC and EHEC strains and to investigate its function and role in pathogenesis. espV was found in 16% of the tested strains. While deletion of espV from C. rodentium did not affect colonization dynamics or fitness in mixed infections, expression of EspV in mammalian cells led to drastic morphological alterations, which were characterized by nuclear condensation, cell rounding, and formation of dendrite-like projections. Expression of EspV in yeast resulted in a dramatic increase in cell size and irreversible growth arrest. Although the role of EspV in infection and its target host cell protein(s) require further investigation, the data point to a novel mechanism by which the T3SS subverts cell signaling.
MeSH term(s)	Amino Acid Sequence ; Animals ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Bacterial Secretion Systems/genetics ; Citrobacter rodentium/metabolism ; Enterohemorrhagic Escherichia coli/genetics ; Enterohemorrhagic Escherichia coli/metabolism ; Enterohemorrhagic Escherichia coli/pathogenicity ; Enteropathogenic Escherichia coli/genetics ; Enteropathogenic Escherichia coli/metabolism ; Enteropathogenic Escherichia coli/pathogenicity ; Escherichia coli Infections/genetics ; Escherichia coli Infections/metabolism ; Eukaryotic Cells/microbiology ; Fluorescent Antibody Technique ; Genes, Bacterial/genetics ; HeLa Cells ; Humans ; Mice ; Microscopy, Electron, Scanning ; Molecular Sequence Data ; Virulence
Chemical Substances	Bacterial Proteins ; Bacterial Secretion Systems
Language	English
Publishing date	2010-12-28
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	218698-6
ISSN	1098-5522 ; 0019-9567
ISSN (online)	1098-5522
ISSN	0019-9567
DOI	10.1128/IAI.01003-10
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Article ; Online: Designed coiled-coil peptides inhibit the type three secretion system of enteropathogenic Escherichia coli.

Larzábal, Mariano / Mercado, Elsa C / Vilte, Daniel A / Salazar-González, Hector / Cataldi, Angel / Navarro-Garcia, Fernando

PloS one

2010 Volume 5, Issue 2, Page(s) e9046

Abstract: Background: Enteropathogenic E. coli (EPEC) and enterohemorrhagic E. coli (EHEC) are two ... pedestals by the same strain in HEp-2 cells and impaired the TTSS-mediated protein translocation ... causing a characteristic attaching and effacing lesion (A/E). The TTSS translocates effector proteins ...

Abstract	Background: Enteropathogenic E. coli (EPEC) and enterohemorrhagic E. coli (EHEC) are two categories of E. coli strains associated with human disease. A major virulence factor of both pathotypes is the expression of a type three secretion system (TTSS), responsible for their ability to adhere to gut mucosa causing a characteristic attaching and effacing lesion (A/E). The TTSS translocates effector proteins directly into the host cell that subvert mammalian cell biochemistry. Methods/principal findings: We examined synthetic peptides designed to inhibit the TTSS. CoilA and CoilB peptides, both representing coiled-coil regions of the translocator protein EspA, and CoilD peptide, corresponding to a coiled-coil region of the needle protein EscF, were effective in inhibiting the TTSS dependent hemolysis of red blood cells by the EPEC E2348/69 strain. CoilA and CoilB peptides also reduced the formation of actin pedestals by the same strain in HEp-2 cells and impaired the TTSS-mediated protein translocation into the epithelial cell. Interestingly, CoilA and CoilB were able to block EspA assembly, destabilizing the TTSS and thereby Tir translocation. This blockage of EspA polymerization by CoilA or CoilB peptides, also inhibited the correct delivery of EspB and EspD as detected by immunoblotting. Interestingly, electron microscopy of bacteria incubated with the CoilA peptide showed a reduction of the length of EspA filaments. Conclusions: Our data indicate that coiled-coil peptides can prevent the assembly and thus the functionality of the TTSS apparatus and suggest that these peptides could provide an attractive tool to block EPEC and EHEC pathogenesis.
MeSH term(s)	Amino Acid Sequence ; Animals ; Bacterial Outer Membrane Proteins/metabolism ; Blotting, Western ; Cell Line, Tumor ; Cytoskeletal Proteins/chemistry ; Cytoskeletal Proteins/metabolism ; Enteropathogenic Escherichia coli/drug effects ; Enteropathogenic Escherichia coli/metabolism ; Erythrocytes/drug effects ; Erythrocytes/microbiology ; Escherichia coli Infections/microbiology ; Escherichia coli Proteins/chemistry ; Escherichia coli Proteins/metabolism ; Escherichia coli Proteins/ultrastructure ; Hemolysis/drug effects ; Humans ; Microbial Viability/drug effects ; Microscopy, Immunoelectron ; Molecular Sequence Data ; Peptides/chemical synthesis ; Peptides/pharmacology ; Protein Transport/drug effects ; Sheep
Chemical Substances	Bacterial Outer Membrane Proteins ; Cytoskeletal Proteins ; EaeB protein, E coli ; EscF protein, E coli ; Escherichia coli Proteins ; EspA protein, E coli ; EspD protein, E coli ; Peptides
Language	English
Publishing date	2010-02-04
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0009046
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Article: Distribution of tccP in clinical enterohemorrhagic and enteropathogenic Escherichia coli isolates.

Garmendia, Junkal / Ren, Zhihong / Tennant, Sharon / Midolli Viera, Monica Aparecida / Chong, Yuwen / Whale, Andrew / Azzopardi, Kristy / Dahan, Sivan / Sircili, Marcelo Palma / Franzolin, Marcia Regina / Trabulsi, Luiz R / Phillips, Alan / Gomes, Tânia A T / Xu, Jianguo / Robins-Browne, Roy / Frankel, Gad

Journal of clinical microbiology

2005 Volume 43, Issue 11, Page(s) 5715–5720

Abstract: Enterohemorrhagic Escherichia coli (EHEC) and enteropathogenic E. coli (EPEC) are diarrheagenic ... pathogens that colonize the gut through the formation of attaching and effacing lesions, which depend ... Recently, two effector proteins, EspJ and TccP, which are encoded by adjacent genes on prophage CP-933U ...

Abstract	Enterohemorrhagic Escherichia coli (EHEC) and enteropathogenic E. coli (EPEC) are diarrheagenic pathogens that colonize the gut through the formation of attaching and effacing lesions, which depend on the translocation of effector proteins via a locus of enterocyte effacement-encoded type III secretion system. Recently, two effector proteins, EspJ and TccP, which are encoded by adjacent genes on prophage CP-933U in EHEC O157:H7, have been identified. TccP consists of a unique N-terminus region and several proline-rich domains. In this project we determined the distribution of tccP in O157:H7, in non-O157 EHEC, and in typical and atypical EPEC isolates. All the EHEC O157:H7 strains tested were tccP(+). Unexpectedly, tccP was also found in non-O157 EHEC, and in typical and atypical EPEC isolates, particularly in strains belonging to serogroups O26 (EHEC), O119 (typical EPEC), and O55 (atypical EPEC). We recorded some variation in the length of tccP, which reflects diversity in the number of the proline-rich repeats. These results show the existence of a class of "attaching and effacing" pathogens which express a combination of EPEC and EHEC virulence determinants.
MeSH term(s)	Animals ; Australia ; Brazil ; China ; Escherichia coli/chemistry ; Escherichia coli/genetics ; Escherichia coli Infections/microbiology ; Escherichia coli Infections/veterinary ; Escherichia coli O157/chemistry ; Escherichia coli O157/genetics ; Escherichia coli Proteins/genetics ; Food Microbiology ; Genetic Variation ; Humans ; Molecular Sequence Data ; United Kingdom
Chemical Substances	Escherichia coli Proteins ; EspJ protein, E coli ; tccP protein, E coli
Language	English
Publishing date	2005-09-29
Publishing country	United States
Document type	Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	390499-4
ISSN	1098-660X ; 0095-1137
ISSN (online)	1098-660X
ISSN	0095-1137
DOI	10.1128/JCM.43.11.5715-5720.2005
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