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IbeR facilitates stress-resistance, invasion and pathogenicity of avian pathogenic Escherichia coli.

Wang S, Bao Y, Meng Q, Xia Y, Zhao Y, Wang Y, Tang F, ZhuGe X, Yu S, Han X, Dai J, Lu C - PLoS ONE (2015)

Bottom Line: Inactivation of ibeR led to attenuated virulence and reduced invasion capacity towards DF-1 cells, brains and cerebrospinal fluid (CSF) in vitro and in vivo.These virulence-related phenotypes were restored by genetic complementation.Quantitative real-time reverse transcription PCR revealed that IbeR controlled expression of stress-resistance genes and virulence genes, which might led to the associated virulence phenotype.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, China; Key Lab of Animal Bacteriology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, 210095, China.

ABSTRACT
Systemic infections by avian pathogenic Escherichia coli (APEC) are economically devastating to poultry industries worldwide. IbeR, located on genomic island GimA, was shown to serve as an RpoS-like regulator in rpoS gene mutation neonatal meningitis E. coli (NMEC) RS218. However, the role of IbeR in pathogenicity of APEC carrying active RpoS has not yet been investigated. We showed that the APEC IbeR could elicit antibodies in infected ducks, suggesting that IbeR might be involved in APEC pathogenicity. To investigate the function of IbeR in APEC pathogenesis, mutant and complementation strains were constructed and characterized. Inactivation of ibeR led to attenuated virulence and reduced invasion capacity towards DF-1 cells, brains and cerebrospinal fluid (CSF) in vitro and in vivo. Bactericidal assays demonstrated that the mutant strain had impaired resistance to environmental stress and specific pathogen-free (SPF) chicken serum. These virulence-related phenotypes were restored by genetic complementation. Quantitative real-time reverse transcription PCR revealed that IbeR controlled expression of stress-resistance genes and virulence genes, which might led to the associated virulence phenotype.

No MeSH data available.


Related in: MedlinePlus

Expression of IbeR by Western blotting.(A) Immunoblotting with anti-IbeR of total cell lysates from different APEC strains. Expression of IbeR was detected in wild-type strain DE205B and complementation strain CΔibeR, but not mutant strains ΔibeR or PΔibeR. Lane M, prestained protein marker; Lane 1, ΔibeR (ibeR negative); Lane 2, DE205B (ibeR positive); Lane 3, CΔibeR (ibeR positive); Lane 4, PΔibeR (ibeR negative). (B) Immunoblotting of purified IbeR protein using anti-DE205B. Incubation with anti-DE205B detected protein bands of the expected size for purified IbeR protein. Lane 1, anti-DE205B.
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pone.0119698.g001: Expression of IbeR by Western blotting.(A) Immunoblotting with anti-IbeR of total cell lysates from different APEC strains. Expression of IbeR was detected in wild-type strain DE205B and complementation strain CΔibeR, but not mutant strains ΔibeR or PΔibeR. Lane M, prestained protein marker; Lane 1, ΔibeR (ibeR negative); Lane 2, DE205B (ibeR positive); Lane 3, CΔibeR (ibeR positive); Lane 4, PΔibeR (ibeR negative). (B) Immunoblotting of purified IbeR protein using anti-DE205B. Incubation with anti-DE205B detected protein bands of the expected size for purified IbeR protein. Lane 1, anti-DE205B.

Mentions: The expression of IbeR in wild-type, mutant, and complementation strains was compared by SDS-PAGE. No differences in protein patterns between the wild-type and mutant strains were detected (data not shown). Immunoblotting was performed with anti-IbeR serum, showing expected protein bands for IbeR from strains DE205B and CΔibeR. However, no IbeR protein was detected from mutant strain ΔibeR and PΔibeR (Fig. 1). These results indicated that IbeR was expressed under laboratory conditions and verified the construction of the ibeR mutant strain.


IbeR facilitates stress-resistance, invasion and pathogenicity of avian pathogenic Escherichia coli.

Wang S, Bao Y, Meng Q, Xia Y, Zhao Y, Wang Y, Tang F, ZhuGe X, Yu S, Han X, Dai J, Lu C - PLoS ONE (2015)

Expression of IbeR by Western blotting.(A) Immunoblotting with anti-IbeR of total cell lysates from different APEC strains. Expression of IbeR was detected in wild-type strain DE205B and complementation strain CΔibeR, but not mutant strains ΔibeR or PΔibeR. Lane M, prestained protein marker; Lane 1, ΔibeR (ibeR negative); Lane 2, DE205B (ibeR positive); Lane 3, CΔibeR (ibeR positive); Lane 4, PΔibeR (ibeR negative). (B) Immunoblotting of purified IbeR protein using anti-DE205B. Incubation with anti-DE205B detected protein bands of the expected size for purified IbeR protein. Lane 1, anti-DE205B.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4359115&req=5

pone.0119698.g001: Expression of IbeR by Western blotting.(A) Immunoblotting with anti-IbeR of total cell lysates from different APEC strains. Expression of IbeR was detected in wild-type strain DE205B and complementation strain CΔibeR, but not mutant strains ΔibeR or PΔibeR. Lane M, prestained protein marker; Lane 1, ΔibeR (ibeR negative); Lane 2, DE205B (ibeR positive); Lane 3, CΔibeR (ibeR positive); Lane 4, PΔibeR (ibeR negative). (B) Immunoblotting of purified IbeR protein using anti-DE205B. Incubation with anti-DE205B detected protein bands of the expected size for purified IbeR protein. Lane 1, anti-DE205B.
Mentions: The expression of IbeR in wild-type, mutant, and complementation strains was compared by SDS-PAGE. No differences in protein patterns between the wild-type and mutant strains were detected (data not shown). Immunoblotting was performed with anti-IbeR serum, showing expected protein bands for IbeR from strains DE205B and CΔibeR. However, no IbeR protein was detected from mutant strain ΔibeR and PΔibeR (Fig. 1). These results indicated that IbeR was expressed under laboratory conditions and verified the construction of the ibeR mutant strain.

Bottom Line: Inactivation of ibeR led to attenuated virulence and reduced invasion capacity towards DF-1 cells, brains and cerebrospinal fluid (CSF) in vitro and in vivo.These virulence-related phenotypes were restored by genetic complementation.Quantitative real-time reverse transcription PCR revealed that IbeR controlled expression of stress-resistance genes and virulence genes, which might led to the associated virulence phenotype.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, China; Key Lab of Animal Bacteriology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, 210095, China.

ABSTRACT
Systemic infections by avian pathogenic Escherichia coli (APEC) are economically devastating to poultry industries worldwide. IbeR, located on genomic island GimA, was shown to serve as an RpoS-like regulator in rpoS gene mutation neonatal meningitis E. coli (NMEC) RS218. However, the role of IbeR in pathogenicity of APEC carrying active RpoS has not yet been investigated. We showed that the APEC IbeR could elicit antibodies in infected ducks, suggesting that IbeR might be involved in APEC pathogenicity. To investigate the function of IbeR in APEC pathogenesis, mutant and complementation strains were constructed and characterized. Inactivation of ibeR led to attenuated virulence and reduced invasion capacity towards DF-1 cells, brains and cerebrospinal fluid (CSF) in vitro and in vivo. Bactericidal assays demonstrated that the mutant strain had impaired resistance to environmental stress and specific pathogen-free (SPF) chicken serum. These virulence-related phenotypes were restored by genetic complementation. Quantitative real-time reverse transcription PCR revealed that IbeR controlled expression of stress-resistance genes and virulence genes, which might led to the associated virulence phenotype.

No MeSH data available.


Related in: MedlinePlus