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GidA, a tRNA Modification Enzyme, Contributes to the Growth, and Virulence of Streptococcus suis Serotype 2.

Gao T, Tan M, Liu W, Zhang C, Zhang T, Zheng L, Zhu J, Li L, Zhou R - Front Cell Infect Microbiol (2016)

Bottom Line: Here, we report a GidA homolog from a Chinese isolate SC-19 of the zoonotic Streptococcus suis serotype 2 (SS2). gidA disruption led to a defective growth, increased capsule thickness, and reduced hemolytic activity.This is consistent with the phenotypes of the mutant.Our findings provide new insight into the regulatory function of GidA in bacterial pathogens.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityWuhan, China; Veterinary Medicine Laboratory, Institute of Animal Husbandry and Veterinary Science, Hubei Academy of Agricultural ScienceWuhan, China; Wuhan Chopper Biology Co., Ltd.Wuhan, China.

ABSTRACT
Glucose-inhibited division protein (GidA), is a tRNA modification enzyme functioning together with MnmE in the addition of a carboxymethylaminomethyl group to position 5 of the anticodon wobble uridine of tRNA. Here, we report a GidA homolog from a Chinese isolate SC-19 of the zoonotic Streptococcus suis serotype 2 (SS2). gidA disruption led to a defective growth, increased capsule thickness, and reduced hemolytic activity. Moreover, the gidA deletion mutant (ΔgidA) displayed reduced mortality and bacterial loads in mice, reduced ability of adhesion to and invasion in epithelial cells, and increased sensitivity to phagocytosis. The iTRAQ analysis identified 372 differentially expressed (182 up- and 190 down-regulated) proteins in ΔgidA and SC-19. Numerous DNA replication, cell division, and virulence associated proteins were downregulated, whereas many capsule synthesis enzymes were upregulated by gidA disruption. This is consistent with the phenotypes of the mutant. Thus, GidA is a translational regulator that plays an important role in the growth, cell division, capsule biosynthesis, and virulence of SS2. Our findings provide new insight into the regulatory function of GidA in bacterial pathogens.

No MeSH data available.


Related in: MedlinePlus

Phagocytosis, adhesion, and invasion assays. (A) Phagocytosis of S. suis by murine macrophages. SC-19 and ΔgidA mutant were incubated with RAW264.7 cells for 30 min at a multiplicity of infection of 10 (MOI = 10:1). Penicillin (100 μg/ml) was then used to kill the extracellular bacteria. The mutant strain ΔgidA showed significantly reduced anti-phagocytosis capability compared with SC-19 (***p < 0.001). (B) The mutant strain ΔgidA showed significantly reduced levels of adherence to HEp-2 cells compared with the = degree of adherence of SC-19 (*p < 0.05). (C) The mutant strain ΔgidA showed significantly reduced levels of invasion of HEp-2 cells compared with that of SC-19 (**p < 0.01).
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Figure 4: Phagocytosis, adhesion, and invasion assays. (A) Phagocytosis of S. suis by murine macrophages. SC-19 and ΔgidA mutant were incubated with RAW264.7 cells for 30 min at a multiplicity of infection of 10 (MOI = 10:1). Penicillin (100 μg/ml) was then used to kill the extracellular bacteria. The mutant strain ΔgidA showed significantly reduced anti-phagocytosis capability compared with SC-19 (***p < 0.001). (B) The mutant strain ΔgidA showed significantly reduced levels of adherence to HEp-2 cells compared with the = degree of adherence of SC-19 (*p < 0.05). (C) The mutant strain ΔgidA showed significantly reduced levels of invasion of HEp-2 cells compared with that of SC-19 (**p < 0.01).

Mentions: To investigate the role of gidA on phagocytosis of S. suis, we performed a phagocytosis assay by using RAW264.7 cells. The numbers of intracellular bacteria for ΔgidA (34,320 ± 3130 CFU/well) were approximately two fold higher than those of SC-19 (18430 ± 821 CFU/well; p < 0.001; Figure 4A). This result indicates that inactivation of gidA can impair the capacity of S.suis to resist phagocytosis by macrophages.


GidA, a tRNA Modification Enzyme, Contributes to the Growth, and Virulence of Streptococcus suis Serotype 2.

Gao T, Tan M, Liu W, Zhang C, Zhang T, Zheng L, Zhu J, Li L, Zhou R - Front Cell Infect Microbiol (2016)

Phagocytosis, adhesion, and invasion assays. (A) Phagocytosis of S. suis by murine macrophages. SC-19 and ΔgidA mutant were incubated with RAW264.7 cells for 30 min at a multiplicity of infection of 10 (MOI = 10:1). Penicillin (100 μg/ml) was then used to kill the extracellular bacteria. The mutant strain ΔgidA showed significantly reduced anti-phagocytosis capability compared with SC-19 (***p < 0.001). (B) The mutant strain ΔgidA showed significantly reduced levels of adherence to HEp-2 cells compared with the = degree of adherence of SC-19 (*p < 0.05). (C) The mutant strain ΔgidA showed significantly reduced levels of invasion of HEp-2 cells compared with that of SC-19 (**p < 0.01).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4835480&req=5

Figure 4: Phagocytosis, adhesion, and invasion assays. (A) Phagocytosis of S. suis by murine macrophages. SC-19 and ΔgidA mutant were incubated with RAW264.7 cells for 30 min at a multiplicity of infection of 10 (MOI = 10:1). Penicillin (100 μg/ml) was then used to kill the extracellular bacteria. The mutant strain ΔgidA showed significantly reduced anti-phagocytosis capability compared with SC-19 (***p < 0.001). (B) The mutant strain ΔgidA showed significantly reduced levels of adherence to HEp-2 cells compared with the = degree of adherence of SC-19 (*p < 0.05). (C) The mutant strain ΔgidA showed significantly reduced levels of invasion of HEp-2 cells compared with that of SC-19 (**p < 0.01).
Mentions: To investigate the role of gidA on phagocytosis of S. suis, we performed a phagocytosis assay by using RAW264.7 cells. The numbers of intracellular bacteria for ΔgidA (34,320 ± 3130 CFU/well) were approximately two fold higher than those of SC-19 (18430 ± 821 CFU/well; p < 0.001; Figure 4A). This result indicates that inactivation of gidA can impair the capacity of S.suis to resist phagocytosis by macrophages.

Bottom Line: Here, we report a GidA homolog from a Chinese isolate SC-19 of the zoonotic Streptococcus suis serotype 2 (SS2). gidA disruption led to a defective growth, increased capsule thickness, and reduced hemolytic activity.This is consistent with the phenotypes of the mutant.Our findings provide new insight into the regulatory function of GidA in bacterial pathogens.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityWuhan, China; Veterinary Medicine Laboratory, Institute of Animal Husbandry and Veterinary Science, Hubei Academy of Agricultural ScienceWuhan, China; Wuhan Chopper Biology Co., Ltd.Wuhan, China.

ABSTRACT
Glucose-inhibited division protein (GidA), is a tRNA modification enzyme functioning together with MnmE in the addition of a carboxymethylaminomethyl group to position 5 of the anticodon wobble uridine of tRNA. Here, we report a GidA homolog from a Chinese isolate SC-19 of the zoonotic Streptococcus suis serotype 2 (SS2). gidA disruption led to a defective growth, increased capsule thickness, and reduced hemolytic activity. Moreover, the gidA deletion mutant (ΔgidA) displayed reduced mortality and bacterial loads in mice, reduced ability of adhesion to and invasion in epithelial cells, and increased sensitivity to phagocytosis. The iTRAQ analysis identified 372 differentially expressed (182 up- and 190 down-regulated) proteins in ΔgidA and SC-19. Numerous DNA replication, cell division, and virulence associated proteins were downregulated, whereas many capsule synthesis enzymes were upregulated by gidA disruption. This is consistent with the phenotypes of the mutant. Thus, GidA is a translational regulator that plays an important role in the growth, cell division, capsule biosynthesis, and virulence of SS2. Our findings provide new insight into the regulatory function of GidA in bacterial pathogens.

No MeSH data available.


Related in: MedlinePlus