CRISPR/Cas9 Screens Reveal Requirements for Host Cell Sulfation and Fucosylation in Bacterial Type III Secretion System-Mediated Cytotoxicity

Carlos J. Blondel, Joseph S. Park, Troy P. Hubbard, Alline R. Pacheco, Carole J. Kuehl, Michael J. Walsh, Brigid M. Davis, Benjamin E. Gewurz, John G. Doench, Matthew K. Waldor

Resultado de la investigación: Article

24 Citas (Scopus)

Resumen

Type III secretion systems (T3SSs) inject bacterial effector proteins into host cells and underlie the virulence of many gram-negative pathogens. Studies have illuminated bacterial factors required for T3SS function, but the required host processes remain largely undefined. We coupled CRISPR/Cas9 genome editing technology with the cytotoxicity of two Vibrio parahaemolyticus T3SSs (T3SS1 and T3SS2) to identify human genome disruptions conferring resistance to T3SS-dependent cytotoxicity. We identity non-overlapping genes required for T3SS1- and T3SS2-mediated cytotoxicity. Genetic ablation of cell surface sulfation reduces bacterial adhesion and thereby alters the kinetics of T3SS1-mediated cytotoxicity. Cell surface fucosylation is required for T3SS2-dependent killing, and genetic inhibition of fucosylation prevents membrane insertion of the T3SS2 translocon complex. These findings reveal the importance of ubiquitous surface modifications for T3SS function, potentially explaining the broad tropism of V. parahaemolyticus, and highlight the utility of genome-wide CRISPR/Cas9 screens to discover processes underlying host-pathogen interactions.

Idioma originalEnglish
Páginas (desde-hasta)226-237
Número de páginas12
PublicaciónCell Host and Microbe
Volumen20
N.º2
DOI
EstadoPublished - 10 ago 2016

Huella dactilar

Clustered Regularly Interspaced Short Palindromic Repeats
Bacterial Adhesion
Vibrio parahaemolyticus
Host-Pathogen Interactions
Tropism
Bacterial Proteins
Human Genome
Virulence
Genome
Technology
Membranes
Genes
Type III Secretion Systems

ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Virology

Citar esto

Blondel, Carlos J. ; Park, Joseph S. ; Hubbard, Troy P. ; Pacheco, Alline R. ; Kuehl, Carole J. ; Walsh, Michael J. ; Davis, Brigid M. ; Gewurz, Benjamin E. ; Doench, John G. ; Waldor, Matthew K. / CRISPR/Cas9 Screens Reveal Requirements for Host Cell Sulfation and Fucosylation in Bacterial Type III Secretion System-Mediated Cytotoxicity. En: Cell Host and Microbe. 2016 ; Vol. 20, N.º 2. pp. 226-237.
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abstract = "Type III secretion systems (T3SSs) inject bacterial effector proteins into host cells and underlie the virulence of many gram-negative pathogens. Studies have illuminated bacterial factors required for T3SS function, but the required host processes remain largely undefined. We coupled CRISPR/Cas9 genome editing technology with the cytotoxicity of two Vibrio parahaemolyticus T3SSs (T3SS1 and T3SS2) to identify human genome disruptions conferring resistance to T3SS-dependent cytotoxicity. We identity non-overlapping genes required for T3SS1- and T3SS2-mediated cytotoxicity. Genetic ablation of cell surface sulfation reduces bacterial adhesion and thereby alters the kinetics of T3SS1-mediated cytotoxicity. Cell surface fucosylation is required for T3SS2-dependent killing, and genetic inhibition of fucosylation prevents membrane insertion of the T3SS2 translocon complex. These findings reveal the importance of ubiquitous surface modifications for T3SS function, potentially explaining the broad tropism of V. parahaemolyticus, and highlight the utility of genome-wide CRISPR/Cas9 screens to discover processes underlying host-pathogen interactions.",
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Blondel, CJ, Park, JS, Hubbard, TP, Pacheco, AR, Kuehl, CJ, Walsh, MJ, Davis, BM, Gewurz, BE, Doench, JG & Waldor, MK 2016, 'CRISPR/Cas9 Screens Reveal Requirements for Host Cell Sulfation and Fucosylation in Bacterial Type III Secretion System-Mediated Cytotoxicity', Cell Host and Microbe, vol. 20, n.º 2, pp. 226-237. https://doi.org/10.1016/j.chom.2016.06.010

CRISPR/Cas9 Screens Reveal Requirements for Host Cell Sulfation and Fucosylation in Bacterial Type III Secretion System-Mediated Cytotoxicity. / Blondel, Carlos J.; Park, Joseph S.; Hubbard, Troy P.; Pacheco, Alline R.; Kuehl, Carole J.; Walsh, Michael J.; Davis, Brigid M.; Gewurz, Benjamin E.; Doench, John G.; Waldor, Matthew K.

En: Cell Host and Microbe, Vol. 20, N.º 2, 10.08.2016, p. 226-237.

Resultado de la investigación: Article

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T1 - CRISPR/Cas9 Screens Reveal Requirements for Host Cell Sulfation and Fucosylation in Bacterial Type III Secretion System-Mediated Cytotoxicity

AU - Blondel, Carlos J.

AU - Park, Joseph S.

AU - Hubbard, Troy P.

AU - Pacheco, Alline R.

AU - Kuehl, Carole J.

AU - Walsh, Michael J.

AU - Davis, Brigid M.

AU - Gewurz, Benjamin E.

AU - Doench, John G.

AU - Waldor, Matthew K.

PY - 2016/8/10

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N2 - Type III secretion systems (T3SSs) inject bacterial effector proteins into host cells and underlie the virulence of many gram-negative pathogens. Studies have illuminated bacterial factors required for T3SS function, but the required host processes remain largely undefined. We coupled CRISPR/Cas9 genome editing technology with the cytotoxicity of two Vibrio parahaemolyticus T3SSs (T3SS1 and T3SS2) to identify human genome disruptions conferring resistance to T3SS-dependent cytotoxicity. We identity non-overlapping genes required for T3SS1- and T3SS2-mediated cytotoxicity. Genetic ablation of cell surface sulfation reduces bacterial adhesion and thereby alters the kinetics of T3SS1-mediated cytotoxicity. Cell surface fucosylation is required for T3SS2-dependent killing, and genetic inhibition of fucosylation prevents membrane insertion of the T3SS2 translocon complex. These findings reveal the importance of ubiquitous surface modifications for T3SS function, potentially explaining the broad tropism of V. parahaemolyticus, and highlight the utility of genome-wide CRISPR/Cas9 screens to discover processes underlying host-pathogen interactions.

AB - Type III secretion systems (T3SSs) inject bacterial effector proteins into host cells and underlie the virulence of many gram-negative pathogens. Studies have illuminated bacterial factors required for T3SS function, but the required host processes remain largely undefined. We coupled CRISPR/Cas9 genome editing technology with the cytotoxicity of two Vibrio parahaemolyticus T3SSs (T3SS1 and T3SS2) to identify human genome disruptions conferring resistance to T3SS-dependent cytotoxicity. We identity non-overlapping genes required for T3SS1- and T3SS2-mediated cytotoxicity. Genetic ablation of cell surface sulfation reduces bacterial adhesion and thereby alters the kinetics of T3SS1-mediated cytotoxicity. Cell surface fucosylation is required for T3SS2-dependent killing, and genetic inhibition of fucosylation prevents membrane insertion of the T3SS2 translocon complex. These findings reveal the importance of ubiquitous surface modifications for T3SS function, potentially explaining the broad tropism of V. parahaemolyticus, and highlight the utility of genome-wide CRISPR/Cas9 screens to discover processes underlying host-pathogen interactions.

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