Identification and characterization of NleI, a new non-LEE-encoded effector of enteropathogenic Escherichia coli (EPEC)

Mo Li; Ilan Rosenshine; Hong Bing Yu; Chen Nadler; Erez Mills; Choy Leong Hew; Ka Yin Leung

doi:10.1016/j.micinf.2006.09.006

Identification and characterization of NleI, a new non-LEE-encoded effector of enteropathogenic Escherichia coli (EPEC)

Mo Li, Ilan Rosenshine, Hong Bing Yu, Chen Nadler, Erez Mills, Choy Leong Hew, Ka Yin Leung^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

26 Scopus citations

Abstract

Enteropathogenic Escherichia coli (EPEC), a major gastrointestinal pathogen, causes infantile diarrhea in many developing countries. EPEC is an attaching and effacing (A/E) pathogen that utilizes a LEE-encoded type III secretion system (TTSS) to deliver effector proteins into host cells. These effectors have been identified as potential virulence factors in A/E pathogens including EPEC, enterohemorrhagic E. coli (EHEC) and Citrobacter rodentium (CR). We used a proteomics approach to identify a new non-LEE-encoded effector, NleI, from the EPEC sepL and sepD mutants. The nleI gene, located in a prophage-associated island with nleBCD, is also present in EHEC and CR but not in E. coli K-12. In EPEC, the transcription of nleI was increased upon sepD inactivation but remained unaffected in ler and sepL mutants. We demonstrated that NleI is secreted and translocated into HeLa cells in a TTSS-dependent manner and that the CesT chaperone is required for efficient NleI translocation. Nevertheless, under overexpression conditions, the first 20 amino acids of NleI are sufficient to support both secretion and translocation. After translocation, NleI can be detected in the cytoplasmic and membrane of HeLa cells.

Original language	English
Pages (from-to)	2890-2898
Number of pages	9
Journal	Microbes and Infection
Volume	8
Issue number	14-15
DOIs	https://doi.org/10.1016/j.micinf.2006.09.006
State	Published - Nov 2006

Bibliographical note

Funding Information:
We are grateful to the National University of Singapore and Israeli Academy of Science and Humanities for providing research grants for this work. We extend our thanks to B.B. Finlay for providing us with CR strain DBS100, plasmid pRE112, anti-Tir monoclonal antibody, and the use of microscope facility, to W. Deng, V.A. Garcia-Angulo and J.L. Puente for scientific discussion, and to P. Tang and S. Joshi for critical reading of the manuscript.

Keywords

Effector
Enteropathogenic E. coli
NleI
Protein translocation
Proteomics
Type III secretion system

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10.1016/j.micinf.2006.09.006

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title = "Identification and characterization of NleI, a new non-LEE-encoded effector of enteropathogenic Escherichia coli (EPEC)",

abstract = "Enteropathogenic Escherichia coli (EPEC), a major gastrointestinal pathogen, causes infantile diarrhea in many developing countries. EPEC is an attaching and effacing (A/E) pathogen that utilizes a LEE-encoded type III secretion system (TTSS) to deliver effector proteins into host cells. These effectors have been identified as potential virulence factors in A/E pathogens including EPEC, enterohemorrhagic E. coli (EHEC) and Citrobacter rodentium (CR). We used a proteomics approach to identify a new non-LEE-encoded effector, NleI, from the EPEC sepL and sepD mutants. The nleI gene, located in a prophage-associated island with nleBCD, is also present in EHEC and CR but not in E. coli K-12. In EPEC, the transcription of nleI was increased upon sepD inactivation but remained unaffected in ler and sepL mutants. We demonstrated that NleI is secreted and translocated into HeLa cells in a TTSS-dependent manner and that the CesT chaperone is required for efficient NleI translocation. Nevertheless, under overexpression conditions, the first 20 amino acids of NleI are sufficient to support both secretion and translocation. After translocation, NleI can be detected in the cytoplasmic and membrane of HeLa cells.",

keywords = "Effector, Enteropathogenic E. coli, NleI, Protein translocation, Proteomics, Type III secretion system",

author = "Mo Li and Ilan Rosenshine and Yu, {Hong Bing} and Chen Nadler and Erez Mills and Hew, {Choy Leong} and Leung, {Ka Yin}",

note = "Funding Information: We are grateful to the National University of Singapore and Israeli Academy of Science and Humanities for providing research grants for this work. We extend our thanks to B.B. Finlay for providing us with CR strain DBS100, plasmid pRE112, anti-Tir monoclonal antibody, and the use of microscope facility, to W. Deng, V.A. Garcia-Angulo and J.L. Puente for scientific discussion, and to P. Tang and S. Joshi for critical reading of the manuscript. ",

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AU - Mills, Erez

AU - Hew, Choy Leong

AU - Leung, Ka Yin

N1 - Funding Information: We are grateful to the National University of Singapore and Israeli Academy of Science and Humanities for providing research grants for this work. We extend our thanks to B.B. Finlay for providing us with CR strain DBS100, plasmid pRE112, anti-Tir monoclonal antibody, and the use of microscope facility, to W. Deng, V.A. Garcia-Angulo and J.L. Puente for scientific discussion, and to P. Tang and S. Joshi for critical reading of the manuscript.

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N2 - Enteropathogenic Escherichia coli (EPEC), a major gastrointestinal pathogen, causes infantile diarrhea in many developing countries. EPEC is an attaching and effacing (A/E) pathogen that utilizes a LEE-encoded type III secretion system (TTSS) to deliver effector proteins into host cells. These effectors have been identified as potential virulence factors in A/E pathogens including EPEC, enterohemorrhagic E. coli (EHEC) and Citrobacter rodentium (CR). We used a proteomics approach to identify a new non-LEE-encoded effector, NleI, from the EPEC sepL and sepD mutants. The nleI gene, located in a prophage-associated island with nleBCD, is also present in EHEC and CR but not in E. coli K-12. In EPEC, the transcription of nleI was increased upon sepD inactivation but remained unaffected in ler and sepL mutants. We demonstrated that NleI is secreted and translocated into HeLa cells in a TTSS-dependent manner and that the CesT chaperone is required for efficient NleI translocation. Nevertheless, under overexpression conditions, the first 20 amino acids of NleI are sufficient to support both secretion and translocation. After translocation, NleI can be detected in the cytoplasmic and membrane of HeLa cells.

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JF - Microbes and Infection

IS - 14-15

ER -

Identification and characterization of NleI, a new non-LEE-encoded effector of enteropathogenic Escherichia coli (EPEC)

Abstract

Bibliographical note

Keywords

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