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Identification and characterization of the chromosomal yefM-yoeB toxin-antitoxin system of Streptococcus suis.

Zheng C, Xu J, Ren S, Li J, Xia M, Chen H, Bei W - Sci Rep (2015)

Bottom Line: Overproduction of S. suis YoeB toxin inhibited the growth of E. coli, and the toxicity of S. suis YoeB could be alleviated by the antitoxin YefM from S. suis and Streptococcus pneumoniae, but not by E. coli YefM.In a murine infection model, deletion of the yefM-yoeB locus had no effect on the virulence of S. suis serotype 2.Collectively, our data suggested that the yefM-yoeB locus of S. suis is an active TA system without the involvement of virulence.

View Article: PubMed Central - PubMed

Affiliation: 1] State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei, 430070, China [2] Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, Hubei, 430070, China.

ABSTRACT
Toxin-antitoxin (TA) systems are widely prevalent in the genomes of bacteria and archaea. These modules have been identified in Escherichia coli and various other bacteria. However, their presence in the genome of Streptococcus suis, an important zoonotic pathogen, has received little attention. In this study, we describe the identification and characterization of a type II TA system, comprising the chromosomal yefM-yoeB locus of S. suis. The yefM-yoeB locus is present in the genome of most serotypes of S. suis. Overproduction of S. suis YoeB toxin inhibited the growth of E. coli, and the toxicity of S. suis YoeB could be alleviated by the antitoxin YefM from S. suis and Streptococcus pneumoniae, but not by E. coli YefM. More importantly, introduction of the S. suis yefM-yoeB system into E. coli could affect cell growth. In a murine infection model, deletion of the yefM-yoeB locus had no effect on the virulence of S. suis serotype 2. Collectively, our data suggested that the yefM-yoeB locus of S. suis is an active TA system without the involvement of virulence.

No MeSH data available.


Related in: MedlinePlus

Effect of introduction of the S. suis yefM-yoeB system into E. coli.(a) Growth curves of E. coli strains carrying the pET30a-yefM-yoeB or pET-30a plasmid. Cells were grown in LB-kanamycin. (b) Growth curves of E. coli strains carrying the pSET2-yefM-yoeB or pSET2 plasmid. Cells were grown in LB-spectinomycin. (c) Growth curves of E. coli strains carrying the pBADhisA-yefM-yoeB or pBADhisA plasmid. Cells were grown in LB medium-ampicillin. (d) Growth curves of E. coli BL21(DE3) strain carrying the pET30a-yefM-yoeB or pET-30a plasmid. Cells were grown in LB-kanamycin and 1 mM IPTG. The data shown are averages with standard deviations for the results from three independent experiments.
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f5: Effect of introduction of the S. suis yefM-yoeB system into E. coli.(a) Growth curves of E. coli strains carrying the pET30a-yefM-yoeB or pET-30a plasmid. Cells were grown in LB-kanamycin. (b) Growth curves of E. coli strains carrying the pSET2-yefM-yoeB or pSET2 plasmid. Cells were grown in LB-spectinomycin. (c) Growth curves of E. coli strains carrying the pBADhisA-yefM-yoeB or pBADhisA plasmid. Cells were grown in LB medium-ampicillin. (d) Growth curves of E. coli BL21(DE3) strain carrying the pET30a-yefM-yoeB or pET-30a plasmid. Cells were grown in LB-kanamycin and 1 mM IPTG. The data shown are averages with standard deviations for the results from three independent experiments.

Mentions: It seemed likely that introduction of the S. suis yefM-yoeB system into E. coli affects cell growth, since E. coli Top 10 cells carrying the pBADhisA-yefM-yoeB plasmid showed considerable growth inhibition under both repressed and inductive conditions. To test the hypothesis, the yefM and yoeB genes were cloned together into the pET-30a expression plasmid. When introduction of the pET30a-yefM-yoeB and pET-30a plasmids into E. coli Trans5α and Top10 strains, cells carrying pET30a-yefM-yoeB showed an obvious growth defect compared with cells carrying pET-30a (Fig. 5a). However, when introduction of the two plasmids into E. coli BL21(DE3) strain, no major difference in growth was found (Fig. 5a).


Identification and characterization of the chromosomal yefM-yoeB toxin-antitoxin system of Streptococcus suis.

Zheng C, Xu J, Ren S, Li J, Xia M, Chen H, Bei W - Sci Rep (2015)

Effect of introduction of the S. suis yefM-yoeB system into E. coli.(a) Growth curves of E. coli strains carrying the pET30a-yefM-yoeB or pET-30a plasmid. Cells were grown in LB-kanamycin. (b) Growth curves of E. coli strains carrying the pSET2-yefM-yoeB or pSET2 plasmid. Cells were grown in LB-spectinomycin. (c) Growth curves of E. coli strains carrying the pBADhisA-yefM-yoeB or pBADhisA plasmid. Cells were grown in LB medium-ampicillin. (d) Growth curves of E. coli BL21(DE3) strain carrying the pET30a-yefM-yoeB or pET-30a plasmid. Cells were grown in LB-kanamycin and 1 mM IPTG. The data shown are averages with standard deviations for the results from three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4536659&req=5

f5: Effect of introduction of the S. suis yefM-yoeB system into E. coli.(a) Growth curves of E. coli strains carrying the pET30a-yefM-yoeB or pET-30a plasmid. Cells were grown in LB-kanamycin. (b) Growth curves of E. coli strains carrying the pSET2-yefM-yoeB or pSET2 plasmid. Cells were grown in LB-spectinomycin. (c) Growth curves of E. coli strains carrying the pBADhisA-yefM-yoeB or pBADhisA plasmid. Cells were grown in LB medium-ampicillin. (d) Growth curves of E. coli BL21(DE3) strain carrying the pET30a-yefM-yoeB or pET-30a plasmid. Cells were grown in LB-kanamycin and 1 mM IPTG. The data shown are averages with standard deviations for the results from three independent experiments.
Mentions: It seemed likely that introduction of the S. suis yefM-yoeB system into E. coli affects cell growth, since E. coli Top 10 cells carrying the pBADhisA-yefM-yoeB plasmid showed considerable growth inhibition under both repressed and inductive conditions. To test the hypothesis, the yefM and yoeB genes were cloned together into the pET-30a expression plasmid. When introduction of the pET30a-yefM-yoeB and pET-30a plasmids into E. coli Trans5α and Top10 strains, cells carrying pET30a-yefM-yoeB showed an obvious growth defect compared with cells carrying pET-30a (Fig. 5a). However, when introduction of the two plasmids into E. coli BL21(DE3) strain, no major difference in growth was found (Fig. 5a).

Bottom Line: Overproduction of S. suis YoeB toxin inhibited the growth of E. coli, and the toxicity of S. suis YoeB could be alleviated by the antitoxin YefM from S. suis and Streptococcus pneumoniae, but not by E. coli YefM.In a murine infection model, deletion of the yefM-yoeB locus had no effect on the virulence of S. suis serotype 2.Collectively, our data suggested that the yefM-yoeB locus of S. suis is an active TA system without the involvement of virulence.

View Article: PubMed Central - PubMed

Affiliation: 1] State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei, 430070, China [2] Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, Hubei, 430070, China.

ABSTRACT
Toxin-antitoxin (TA) systems are widely prevalent in the genomes of bacteria and archaea. These modules have been identified in Escherichia coli and various other bacteria. However, their presence in the genome of Streptococcus suis, an important zoonotic pathogen, has received little attention. In this study, we describe the identification and characterization of a type II TA system, comprising the chromosomal yefM-yoeB locus of S. suis. The yefM-yoeB locus is present in the genome of most serotypes of S. suis. Overproduction of S. suis YoeB toxin inhibited the growth of E. coli, and the toxicity of S. suis YoeB could be alleviated by the antitoxin YefM from S. suis and Streptococcus pneumoniae, but not by E. coli YefM. More importantly, introduction of the S. suis yefM-yoeB system into E. coli could affect cell growth. In a murine infection model, deletion of the yefM-yoeB locus had no effect on the virulence of S. suis serotype 2. Collectively, our data suggested that the yefM-yoeB locus of S. suis is an active TA system without the involvement of virulence.

No MeSH data available.


Related in: MedlinePlus