The type VI secretion system encoded in salmonella pathogenicity island 19 is required for Salmonella enterica serotype gallinarum survival within infected macrophages

Carlos J. Blondel, Juan C. Jiménez, Lorenzo E. Leiva, Sergio A. Álvarez, Bernardo I. Pinto, Francisca Contreras, David Pezoa, Carlos A. Santiviago, Inés Contreras

Resultado de la investigación: Article

28 Citas (Scopus)

Resumen

Salmonella enterica serotype Gallinarum is the causative agent of fowl typhoid, a disease characterized by high morbidity and mortality that causes major economic losses in poultry production. We have reported that S. Gallinarum harbors a type VI secretion system (T6SS) encoded in Salmonella pathogenicity island 19 (SPI-19) that is required for efficient colonization of chicks. In the present study, we aimed to characterize the SPI-19 T6SS functionality and to investigate the mechanisms behind the phenotypes previously observed in vivo. Expression analyses revealed that SPI-19 T6SS core components are expressed and produced under in vitro bacterial growth conditions. However, secretion of the structural/secreted components Hcp1, Hcp2, and VgrG to the culture medium could not be determined, suggesting that additional signals are required for T6SS-dependent secretion of these proteins. In vitro bacterial competition assays failed to demonstrate a role for SPI-19 T6SS in interbacterial killing. In contrast, cell culture experiments with murine and avian macrophages (RAW264.7 and HD11, respectively) revealed production of a green fluorescent protein-tagged version of VgrG soon after Salmonella uptake. Furthermore, infection of RAW264.7 and HD11 macrophages with deletion mutants of SPI-19 or strains with genes encoding specific T6SS core components (clpV and vgrG) revealed that SPI-19 T6SS contributes to S. Gallinarum survival within macrophages at 20 h postuptake. SPI-19 T6SS function was not linked to Salmonella-induced cytotoxicity or cell death of infected macrophages, as has been described for other T6SS. Our data indicate that SPI-19 T6SS corresponds to a novel tool used by Salmonella to survive within host cells.

Idioma originalEnglish
Páginas (desde-hasta)1207-1220
Número de páginas14
PublicaciónInfection and Immunity
Volumen81
N.º4
DOI
EstadoPublished - 1 abr 2013

Huella dactilar

Genomic Islands
Salmonella enterica
Salmonella
Macrophages
Type VI Secretion Systems
Serogroup
Typhoid Fever
Poultry
Green Fluorescent Proteins
Culture Media
Cell Death
Cell Culture Techniques
Economics

ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Immunology
  • Infectious Diseases

Citar esto

Blondel, Carlos J. ; Jiménez, Juan C. ; Leiva, Lorenzo E. ; Álvarez, Sergio A. ; Pinto, Bernardo I. ; Contreras, Francisca ; Pezoa, David ; Santiviago, Carlos A. ; Contreras, Inés. / The type VI secretion system encoded in salmonella pathogenicity island 19 is required for Salmonella enterica serotype gallinarum survival within infected macrophages. En: Infection and Immunity. 2013 ; Vol. 81, N.º 4. pp. 1207-1220.
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title = "The type VI secretion system encoded in salmonella pathogenicity island 19 is required for Salmonella enterica serotype gallinarum survival within infected macrophages",
abstract = "Salmonella enterica serotype Gallinarum is the causative agent of fowl typhoid, a disease characterized by high morbidity and mortality that causes major economic losses in poultry production. We have reported that S. Gallinarum harbors a type VI secretion system (T6SS) encoded in Salmonella pathogenicity island 19 (SPI-19) that is required for efficient colonization of chicks. In the present study, we aimed to characterize the SPI-19 T6SS functionality and to investigate the mechanisms behind the phenotypes previously observed in vivo. Expression analyses revealed that SPI-19 T6SS core components are expressed and produced under in vitro bacterial growth conditions. However, secretion of the structural/secreted components Hcp1, Hcp2, and VgrG to the culture medium could not be determined, suggesting that additional signals are required for T6SS-dependent secretion of these proteins. In vitro bacterial competition assays failed to demonstrate a role for SPI-19 T6SS in interbacterial killing. In contrast, cell culture experiments with murine and avian macrophages (RAW264.7 and HD11, respectively) revealed production of a green fluorescent protein-tagged version of VgrG soon after Salmonella uptake. Furthermore, infection of RAW264.7 and HD11 macrophages with deletion mutants of SPI-19 or strains with genes encoding specific T6SS core components (clpV and vgrG) revealed that SPI-19 T6SS contributes to S. Gallinarum survival within macrophages at 20 h postuptake. SPI-19 T6SS function was not linked to Salmonella-induced cytotoxicity or cell death of infected macrophages, as has been described for other T6SS. Our data indicate that SPI-19 T6SS corresponds to a novel tool used by Salmonella to survive within host cells.",
author = "Blondel, {Carlos J.} and Jim{\'e}nez, {Juan C.} and Leiva, {Lorenzo E.} and {\'A}lvarez, {Sergio A.} and Pinto, {Bernardo I.} and Francisca Contreras and David Pezoa and Santiviago, {Carlos A.} and In{\'e}s Contreras",
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The type VI secretion system encoded in salmonella pathogenicity island 19 is required for Salmonella enterica serotype gallinarum survival within infected macrophages. / Blondel, Carlos J.; Jiménez, Juan C.; Leiva, Lorenzo E.; Álvarez, Sergio A.; Pinto, Bernardo I.; Contreras, Francisca; Pezoa, David; Santiviago, Carlos A.; Contreras, Inés.

En: Infection and Immunity, Vol. 81, N.º 4, 01.04.2013, p. 1207-1220.

Resultado de la investigación: Article

TY - JOUR

T1 - The type VI secretion system encoded in salmonella pathogenicity island 19 is required for Salmonella enterica serotype gallinarum survival within infected macrophages

AU - Blondel, Carlos J.

AU - Jiménez, Juan C.

AU - Leiva, Lorenzo E.

AU - Álvarez, Sergio A.

AU - Pinto, Bernardo I.

AU - Contreras, Francisca

AU - Pezoa, David

AU - Santiviago, Carlos A.

AU - Contreras, Inés

PY - 2013/4/1

Y1 - 2013/4/1

N2 - Salmonella enterica serotype Gallinarum is the causative agent of fowl typhoid, a disease characterized by high morbidity and mortality that causes major economic losses in poultry production. We have reported that S. Gallinarum harbors a type VI secretion system (T6SS) encoded in Salmonella pathogenicity island 19 (SPI-19) that is required for efficient colonization of chicks. In the present study, we aimed to characterize the SPI-19 T6SS functionality and to investigate the mechanisms behind the phenotypes previously observed in vivo. Expression analyses revealed that SPI-19 T6SS core components are expressed and produced under in vitro bacterial growth conditions. However, secretion of the structural/secreted components Hcp1, Hcp2, and VgrG to the culture medium could not be determined, suggesting that additional signals are required for T6SS-dependent secretion of these proteins. In vitro bacterial competition assays failed to demonstrate a role for SPI-19 T6SS in interbacterial killing. In contrast, cell culture experiments with murine and avian macrophages (RAW264.7 and HD11, respectively) revealed production of a green fluorescent protein-tagged version of VgrG soon after Salmonella uptake. Furthermore, infection of RAW264.7 and HD11 macrophages with deletion mutants of SPI-19 or strains with genes encoding specific T6SS core components (clpV and vgrG) revealed that SPI-19 T6SS contributes to S. Gallinarum survival within macrophages at 20 h postuptake. SPI-19 T6SS function was not linked to Salmonella-induced cytotoxicity or cell death of infected macrophages, as has been described for other T6SS. Our data indicate that SPI-19 T6SS corresponds to a novel tool used by Salmonella to survive within host cells.

AB - Salmonella enterica serotype Gallinarum is the causative agent of fowl typhoid, a disease characterized by high morbidity and mortality that causes major economic losses in poultry production. We have reported that S. Gallinarum harbors a type VI secretion system (T6SS) encoded in Salmonella pathogenicity island 19 (SPI-19) that is required for efficient colonization of chicks. In the present study, we aimed to characterize the SPI-19 T6SS functionality and to investigate the mechanisms behind the phenotypes previously observed in vivo. Expression analyses revealed that SPI-19 T6SS core components are expressed and produced under in vitro bacterial growth conditions. However, secretion of the structural/secreted components Hcp1, Hcp2, and VgrG to the culture medium could not be determined, suggesting that additional signals are required for T6SS-dependent secretion of these proteins. In vitro bacterial competition assays failed to demonstrate a role for SPI-19 T6SS in interbacterial killing. In contrast, cell culture experiments with murine and avian macrophages (RAW264.7 and HD11, respectively) revealed production of a green fluorescent protein-tagged version of VgrG soon after Salmonella uptake. Furthermore, infection of RAW264.7 and HD11 macrophages with deletion mutants of SPI-19 or strains with genes encoding specific T6SS core components (clpV and vgrG) revealed that SPI-19 T6SS contributes to S. Gallinarum survival within macrophages at 20 h postuptake. SPI-19 T6SS function was not linked to Salmonella-induced cytotoxicity or cell death of infected macrophages, as has been described for other T6SS. Our data indicate that SPI-19 T6SS corresponds to a novel tool used by Salmonella to survive within host cells.

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DO - 10.1128/IAI.01165-12

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