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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

yqiC is essential for virulence in mice. BALB/c mice were orally infected with 1 × 105 CFU of wild-type S. Typhimurium ATCC 14028, 14028 ΔyqiC::CAT or 14028 ΔyqiC::CAT + pBBR-yqiC. The survival of infected mice over time is shown.
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Figure 7: yqiC is essential for virulence in mice. BALB/c mice were orally infected with 1 × 105 CFU of wild-type S. Typhimurium ATCC 14028, 14028 ΔyqiC::CAT or 14028 ΔyqiC::CAT + pBBR-yqiC. The survival of infected mice over time is shown.

Mentions: In spite of the clear effect of the yqiC mutant strain on growth at 37°C, we did not observe any defect in colonizing and surviving inside in vitro cultured eukaryotic cells grown at 37°C. Thus, we evaluated the virulence of the yqiC mutant in the murine model. To this aim, we performed oral infections with S. Typhimurium ATCC 14028, 14028 ΔyqiC::CAT and 14028 ΔyqiC::CAT trans-complemented with yqiC in BALB/c mice. As illustrated in Figure 7, no survival was registered by day 20 in mice inoculated with WT strain. However, the yqiC mutant showed complete attenuation in virulence, as all mice infected with this strain survived along the 30-day period of the experiment. The yqiC gene provided in trans fully complemented the 14028 ΔyqiC::CAT phenotype, causing 100% mice death by day 19. In addition, we determined the LD50 of S. Typhimurium ATCC 14028 and 14028 ΔyqiC::CAT in mice inoculated intraperitoneally as described in Materials and methods. A dramatic increase in the LD50 was observed in the yqiC defective strain (>5 × 105 CFU), as compared with the WT (10-100 CFU) (Table 1). Together, these results clearly show that YqiC is required for S. Typhimurium virulence in the murine infection model.


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)

yqiC is essential for virulence in mice. BALB/c mice were orally infected with 1 × 105 CFU of wild-type S. Typhimurium ATCC 14028, 14028 ΔyqiC::CAT or 14028 ΔyqiC::CAT + pBBR-yqiC. The survival of infected mice over time is shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 7: yqiC is essential for virulence in mice. BALB/c mice were orally infected with 1 × 105 CFU of wild-type S. Typhimurium ATCC 14028, 14028 ΔyqiC::CAT or 14028 ΔyqiC::CAT + pBBR-yqiC. The survival of infected mice over time is shown.
Mentions: In spite of the clear effect of the yqiC mutant strain on growth at 37°C, we did not observe any defect in colonizing and surviving inside in vitro cultured eukaryotic cells grown at 37°C. Thus, we evaluated the virulence of the yqiC mutant in the murine model. To this aim, we performed oral infections with S. Typhimurium ATCC 14028, 14028 ΔyqiC::CAT and 14028 ΔyqiC::CAT trans-complemented with yqiC in BALB/c mice. As illustrated in Figure 7, no survival was registered by day 20 in mice inoculated with WT strain. However, the yqiC mutant showed complete attenuation in virulence, as all mice infected with this strain survived along the 30-day period of the experiment. The yqiC gene provided in trans fully complemented the 14028 ΔyqiC::CAT phenotype, causing 100% mice death by day 19. In addition, we determined the LD50 of S. Typhimurium ATCC 14028 and 14028 ΔyqiC::CAT in mice inoculated intraperitoneally as described in Materials and methods. A dramatic increase in the LD50 was observed in the yqiC defective strain (>5 × 105 CFU), as compared with the WT (10-100 CFU) (Table 1). Together, these results clearly show that YqiC is required for S. Typhimurium virulence in the murine infection model.

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