Transposon-insertion sequencing screens unveil requirements for EHEC growth and intestinal colonization

Alyson R. Warr, Troy P. Hubbard, Diana Munera, Carlos J. Blondel, Pia Abel Zur Wiesch, Sören Abel, Xiaoxue Wang, Brigid M. Davis, Matthew K. Waldor

Resultado de la investigación: Article

Resumen

Enterohemorrhagic Escherichia coli O157:H7 (EHEC) is an important food-borne pathogen that colonizes the colon. Transposon-insertion sequencing (TIS) was used to identify genes required for EHEC and E. coli K-12 growth in vitro and for EHEC growth in vivo in the infant rabbit colon. Surprisingly, many conserved loci contribute to EHEC's but not to K-12's growth in vitro. There was a restrictive bottleneck for EHEC colonization of the rabbit colon, which complicated identification of EHEC genes facilitating growth in vivo. Both a refined version of an existing analytic framework as well as PCA-based analysis were used to compensate for the effects of the infection bottleneck. These analyses confirmed that the EHEC LEE-encoded type III secretion apparatus is required for growth in vivo and revealed that only a few effectors are critical for in vivo fitness. Over 200 mutants not previously associated with EHEC survival/growth in vivo also appeared attenuated in vivo, and a subset of these putative in vivo fitness factors were validated. Some were found to contribute to efficient type-three secretion while others, including tatABC, oxyR, envC, acrAB, and cvpA, promote EHEC resistance to host-derived stresses. cvpA is also required for intestinal growth of several other enteric pathogens, and proved to be required for EHEC, Vibrio cholerae and Vibrio parahaemolyticus resistance to the bile salt deoxycholate, highlighting the important role of this previously uncharacterized protein in pathogen survival. Collectively, our findings provide a comprehensive framework for understanding EHEC growth in the intestine.

Idioma originalEnglish
Número de artículoe1007652
PublicaciónPLoS Pathogens
Volumen15
N.º8
DOI
EstadoPublished - 1 ene 2019

Huella dactilar

Enterohemorrhagic Escherichia coli
Escherichia coli O157
Growth
Colon
Rabbits
Vibrio parahaemolyticus
Passive Cutaneous Anaphylaxis
Deoxycholic Acid
Vibrio cholerae
Bile Acids and Salts
Genes
Intestines

ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Immunology
  • Molecular Biology
  • Genetics
  • Virology

Citar esto

Warr, A. R., Hubbard, T. P., Munera, D., Blondel, C. J., Zur Wiesch, P. A., Abel, S., ... Waldor, M. K. (2019). Transposon-insertion sequencing screens unveil requirements for EHEC growth and intestinal colonization. PLoS Pathogens, 15(8), [e1007652]. https://doi.org/10.1371/journal.ppat.1007652
Warr, Alyson R. ; Hubbard, Troy P. ; Munera, Diana ; Blondel, Carlos J. ; Zur Wiesch, Pia Abel ; Abel, Sören ; Wang, Xiaoxue ; Davis, Brigid M. ; Waldor, Matthew K. / Transposon-insertion sequencing screens unveil requirements for EHEC growth and intestinal colonization. En: PLoS Pathogens. 2019 ; Vol. 15, N.º 8.
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abstract = "Enterohemorrhagic Escherichia coli O157:H7 (EHEC) is an important food-borne pathogen that colonizes the colon. Transposon-insertion sequencing (TIS) was used to identify genes required for EHEC and E. coli K-12 growth in vitro and for EHEC growth in vivo in the infant rabbit colon. Surprisingly, many conserved loci contribute to EHEC's but not to K-12's growth in vitro. There was a restrictive bottleneck for EHEC colonization of the rabbit colon, which complicated identification of EHEC genes facilitating growth in vivo. Both a refined version of an existing analytic framework as well as PCA-based analysis were used to compensate for the effects of the infection bottleneck. These analyses confirmed that the EHEC LEE-encoded type III secretion apparatus is required for growth in vivo and revealed that only a few effectors are critical for in vivo fitness. Over 200 mutants not previously associated with EHEC survival/growth in vivo also appeared attenuated in vivo, and a subset of these putative in vivo fitness factors were validated. Some were found to contribute to efficient type-three secretion while others, including tatABC, oxyR, envC, acrAB, and cvpA, promote EHEC resistance to host-derived stresses. cvpA is also required for intestinal growth of several other enteric pathogens, and proved to be required for EHEC, Vibrio cholerae and Vibrio parahaemolyticus resistance to the bile salt deoxycholate, highlighting the important role of this previously uncharacterized protein in pathogen survival. Collectively, our findings provide a comprehensive framework for understanding EHEC growth in the intestine.",
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Warr, AR, Hubbard, TP, Munera, D, Blondel, CJ, Zur Wiesch, PA, Abel, S, Wang, X, Davis, BM & Waldor, MK 2019, 'Transposon-insertion sequencing screens unveil requirements for EHEC growth and intestinal colonization', PLoS Pathogens, vol. 15, n.º 8, e1007652. https://doi.org/10.1371/journal.ppat.1007652

Transposon-insertion sequencing screens unveil requirements for EHEC growth and intestinal colonization. / Warr, Alyson R.; Hubbard, Troy P.; Munera, Diana; Blondel, Carlos J.; Zur Wiesch, Pia Abel; Abel, Sören; Wang, Xiaoxue; Davis, Brigid M.; Waldor, Matthew K.

En: PLoS Pathogens, Vol. 15, N.º 8, e1007652, 01.01.2019.

Resultado de la investigación: Article

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T1 - Transposon-insertion sequencing screens unveil requirements for EHEC growth and intestinal colonization

AU - Warr, Alyson R.

AU - Hubbard, Troy P.

AU - Munera, Diana

AU - Blondel, Carlos J.

AU - Zur Wiesch, Pia Abel

AU - Abel, Sören

AU - Wang, Xiaoxue

AU - Davis, Brigid M.

AU - Waldor, Matthew K.

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N2 - Enterohemorrhagic Escherichia coli O157:H7 (EHEC) is an important food-borne pathogen that colonizes the colon. Transposon-insertion sequencing (TIS) was used to identify genes required for EHEC and E. coli K-12 growth in vitro and for EHEC growth in vivo in the infant rabbit colon. Surprisingly, many conserved loci contribute to EHEC's but not to K-12's growth in vitro. There was a restrictive bottleneck for EHEC colonization of the rabbit colon, which complicated identification of EHEC genes facilitating growth in vivo. Both a refined version of an existing analytic framework as well as PCA-based analysis were used to compensate for the effects of the infection bottleneck. These analyses confirmed that the EHEC LEE-encoded type III secretion apparatus is required for growth in vivo and revealed that only a few effectors are critical for in vivo fitness. Over 200 mutants not previously associated with EHEC survival/growth in vivo also appeared attenuated in vivo, and a subset of these putative in vivo fitness factors were validated. Some were found to contribute to efficient type-three secretion while others, including tatABC, oxyR, envC, acrAB, and cvpA, promote EHEC resistance to host-derived stresses. cvpA is also required for intestinal growth of several other enteric pathogens, and proved to be required for EHEC, Vibrio cholerae and Vibrio parahaemolyticus resistance to the bile salt deoxycholate, highlighting the important role of this previously uncharacterized protein in pathogen survival. Collectively, our findings provide a comprehensive framework for understanding EHEC growth in the intestine.

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