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YqiC of Salmonella enterica serovar Typhimurium is a membrane fusogenic protein required for mice colonization.

Carrica MC, Craig PO, García-Angulo VA, Aguirre A, García-Véscovi E, Goldbaum FA, Cravero SL - BMC Microbiol. (2011)

Bottom Line: We found that YqiC shares biophysical and biochemical properties with Brucella abortus BMFP, the only previously characterized member of this group, such as a high alpha helix content, a coiled-coil domain involved in trimerization and a membrane fusogenic activity in vitro.In addition, we demonstrated that YqiC localizes at cytoplasmic and membrane subcellular fractions, that a S.This work firstly demonstrates the importance of a COG2960 member for pathogen-host interaction, and suggests a common function conserved among members of this group.

View Article: PubMed Central - HTML - PubMed

Affiliation: Instituto de Biotecnología, CICVyA-INTA Castelar, Los Reseros y Las Cabañas s/n, Buenos Aires, Argentina. mcarrica@leloir.org.ar

ABSTRACT

Background: Salmonella enterica serovar Typhimurium is an intracellular bacterial pathogen which can colonize a variety of hosts, including human, causing syndromes that vary from gastroenteritis and diarrhea to systemic disease.

Results: In this work we present structural information as well as insights into the in vivo function of YqiC, a 99-residue protein of S. Typhimurium, which belongs to the cluster of the orthologous group 2960 (COG2960). We found that YqiC shares biophysical and biochemical properties with Brucella abortus BMFP, the only previously characterized member of this group, such as a high alpha helix content, a coiled-coil domain involved in trimerization and a membrane fusogenic activity in vitro. In addition, we demonstrated that YqiC localizes at cytoplasmic and membrane subcellular fractions, that a S. Typhimurium yqiC deficient strain had a severe attenuation in virulence in the murine model when inoculated both orally and intraperitoneally, and was impaired to replicate at physiological and high temperatures in vitro, although it was still able to invade and replicate inside epithelial and macrophages cell lines.

Conclusion: This work firstly demonstrates the importance of a COG2960 member for pathogen-host interaction, and suggests a common function conserved among members of this group.

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Related in: MedlinePlus

Invasion and intracellular survival of S. Typhimurium strains in cultured cells. S. Typhimurium ATCC 14028 (open bars) and 14028 ΔyqiC::CAT mutant (filled bars) recovered from lysates of J774 murine macrophages (A) or human epithelial HeLa cells (B). The number of viable bacteria from cell lysates was determined 1, 6 and 24 hours post infection as described in Materials and methods. The reported value is the media of duplicates of a representative experiment +/- standard deviation.
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Figure 6: Invasion and intracellular survival of S. Typhimurium strains in cultured cells. S. Typhimurium ATCC 14028 (open bars) and 14028 ΔyqiC::CAT mutant (filled bars) recovered from lysates of J774 murine macrophages (A) or human epithelial HeLa cells (B). The number of viable bacteria from cell lysates was determined 1, 6 and 24 hours post infection as described in Materials and methods. The reported value is the media of duplicates of a representative experiment +/- standard deviation.

Mentions: The pathogenicity of S. Typhimurium is critically dependent on its ability to infect and multiply into eukaryotic cells. We investigated whether the 14028 ΔyqiC::CAT strain was affected in its ability to invade and survive within cultured eukaryotic cells. J774 murine macrophages and HeLa human epithelial cell lines were infected with WT S. Typhimurium and 14028 ΔyqiC::CAT strains. As the 14028 ΔyqiC::CAT strain grows defectively at physiological temperature, all strains were grown at 28°C prior to infection. Infected cells were kept at 37°C and viable intracellular bacteria was determined in cell lysates at 1, 6 and 24 hours after infection. In both cell types, no differences were detected at all time points examined in the CFU recovered from cell lysates infected with the WT or the yqiC mutant strains (Figure 6). This result indicates that the yqiC gene does not contribute to neither Salmonella entry nor intracellular survival in the cell types assayed.


YqiC of Salmonella enterica serovar Typhimurium is a membrane fusogenic protein required for mice colonization.

Carrica MC, Craig PO, García-Angulo VA, Aguirre A, García-Véscovi E, Goldbaum FA, Cravero SL - BMC Microbiol. (2011)

Invasion and intracellular survival of S. Typhimurium strains in cultured cells. S. Typhimurium ATCC 14028 (open bars) and 14028 ΔyqiC::CAT mutant (filled bars) recovered from lysates of J774 murine macrophages (A) or human epithelial HeLa cells (B). The number of viable bacteria from cell lysates was determined 1, 6 and 24 hours post infection as described in Materials and methods. The reported value is the media of duplicates of a representative experiment +/- standard deviation.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3107778&req=5

Figure 6: Invasion and intracellular survival of S. Typhimurium strains in cultured cells. S. Typhimurium ATCC 14028 (open bars) and 14028 ΔyqiC::CAT mutant (filled bars) recovered from lysates of J774 murine macrophages (A) or human epithelial HeLa cells (B). The number of viable bacteria from cell lysates was determined 1, 6 and 24 hours post infection as described in Materials and methods. The reported value is the media of duplicates of a representative experiment +/- standard deviation.
Mentions: The pathogenicity of S. Typhimurium is critically dependent on its ability to infect and multiply into eukaryotic cells. We investigated whether the 14028 ΔyqiC::CAT strain was affected in its ability to invade and survive within cultured eukaryotic cells. J774 murine macrophages and HeLa human epithelial cell lines were infected with WT S. Typhimurium and 14028 ΔyqiC::CAT strains. As the 14028 ΔyqiC::CAT strain grows defectively at physiological temperature, all strains were grown at 28°C prior to infection. Infected cells were kept at 37°C and viable intracellular bacteria was determined in cell lysates at 1, 6 and 24 hours after infection. In both cell types, no differences were detected at all time points examined in the CFU recovered from cell lysates infected with the WT or the yqiC mutant strains (Figure 6). This result indicates that the yqiC gene does not contribute to neither Salmonella entry nor intracellular survival in the cell types assayed.

Bottom Line: We found that YqiC shares biophysical and biochemical properties with Brucella abortus BMFP, the only previously characterized member of this group, such as a high alpha helix content, a coiled-coil domain involved in trimerization and a membrane fusogenic activity in vitro.In addition, we demonstrated that YqiC localizes at cytoplasmic and membrane subcellular fractions, that a S.This work firstly demonstrates the importance of a COG2960 member for pathogen-host interaction, and suggests a common function conserved among members of this group.

View Article: PubMed Central - HTML - PubMed

Affiliation: Instituto de Biotecnología, CICVyA-INTA Castelar, Los Reseros y Las Cabañas s/n, Buenos Aires, Argentina. mcarrica@leloir.org.ar

ABSTRACT

Background: Salmonella enterica serovar Typhimurium is an intracellular bacterial pathogen which can colonize a variety of hosts, including human, causing syndromes that vary from gastroenteritis and diarrhea to systemic disease.

Results: In this work we present structural information as well as insights into the in vivo function of YqiC, a 99-residue protein of S. Typhimurium, which belongs to the cluster of the orthologous group 2960 (COG2960). We found that YqiC shares biophysical and biochemical properties with Brucella abortus BMFP, the only previously characterized member of this group, such as a high alpha helix content, a coiled-coil domain involved in trimerization and a membrane fusogenic activity in vitro. In addition, we demonstrated that YqiC localizes at cytoplasmic and membrane subcellular fractions, that a S. Typhimurium yqiC deficient strain had a severe attenuation in virulence in the murine model when inoculated both orally and intraperitoneally, and was impaired to replicate at physiological and high temperatures in vitro, although it was still able to invade and replicate inside epithelial and macrophages cell lines.

Conclusion: This work firstly demonstrates the importance of a COG2960 member for pathogen-host interaction, and suggests a common function conserved among members of this group.

Show MeSH
Related in: MedlinePlus