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MsmK, an ATPase, Contributes to Utilization of Multiple Carbohydrates and Host Colonization of Streptococcus suis.

Tan MF, Gao T, Liu WQ, Zhang CY, Yang X, Zhu JW, Teng MY, Li L, Zhou R - PLoS ONE (2015)

Bottom Line: Genetic and biochemistry studies revealed that the MsmK was responsible for the utilization of raffinose, melibiose, maltotetraose, glycogen and maltotriose.In infected mice, the msmK-deletion mutant showed significant defects of survival and colonization when compared with its parental and complementary strains.Taken together, MsmK is an ATPase that contributes to multiple carbohydrates utilization and host colonization of S. suis.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China.

ABSTRACT
Acquisition and metabolism of carbohydrates are essential for host colonization and pathogenesis of bacterial pathogens. Different bacteria can uptake different lines of carbohydrates via ABC transporters, in which ATPase subunits energize the transport though ATP hydrolysis. Some ABC transporters possess their own ATPases, while some share a common ATPase. Here we identified MsmK, an ATPase from Streptococcus suis, an emerging zoonotic bacterium causing dead infections in pigs and humans. Genetic and biochemistry studies revealed that the MsmK was responsible for the utilization of raffinose, melibiose, maltotetraose, glycogen and maltotriose. In infected mice, the msmK-deletion mutant showed significant defects of survival and colonization when compared with its parental and complementary strains. Taken together, MsmK is an ATPase that contributes to multiple carbohydrates utilization and host colonization of S. suis. This study gives new insight into our understanding of the carbohydrates utilization and its relationship to the pathogenesis of this zoonotic pathogen.

No MeSH data available.


Related in: MedlinePlus

Western blot analysis of MsmK expression in SC-19 cultured in different carbohydrates.(A) Strain SC-19 was cultured in CDM supplemented with 1% (g/v) diverse carbohydrates to log phase. The supernatant of cell lysate was disposed for immunoblot analysis with MsmK or Enolase polyclonal antibodies. Enolase was used as an internal reference. (B) The blots were scanned, and the resultant graphics were quantified with ImageJ2x. Fold change is represented as a comparison to cells grown in glucose, and the expression levels of Enolase as internal references. Data are shown as means ± SEM from at least three independent experiments. Statistical significance was tested by a Two-way ANOVA test (ns, P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001).
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pone.0130792.g005: Western blot analysis of MsmK expression in SC-19 cultured in different carbohydrates.(A) Strain SC-19 was cultured in CDM supplemented with 1% (g/v) diverse carbohydrates to log phase. The supernatant of cell lysate was disposed for immunoblot analysis with MsmK or Enolase polyclonal antibodies. Enolase was used as an internal reference. (B) The blots were scanned, and the resultant graphics were quantified with ImageJ2x. Fold change is represented as a comparison to cells grown in glucose, and the expression levels of Enolase as internal references. Data are shown as means ± SEM from at least three independent experiments. Statistical significance was tested by a Two-way ANOVA test (ns, P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001).

Mentions: Since MsmK is suspected to energize the MsmEFG and MalXCD transporters in S. suis for multiple carbohydrates transport, it was supposed that the levels of MsmK expressed in CDM with diverse carbohydrates aforementioned would be induced in some degree. To obtain the relative expression levels of MsmK, SC-19 was cultured in CDM supplemented with different carbohydrates as sole carbon source. MsmK expression was evaluated by Western blot (Fig 5A). The experimental images (Fig 5A) were quantified by using ImageJ2x software. The amount of MsmK expressed in SC-19 cultured in CDM with glucose was defined as one fold. As expected, expression of MsmK increased on different levels (Fig 5B). According to the result, expression of MsmK in 1% melibiose, glycogen and raffinose were significantly increased (p ≤ 0.01). In contrast, levels of MsmK in 1% maltotriose and maltotraose had fewer changes, but still higher than that in 1% glucose or maltose. MsmK expression induced by these carbohydrates further supports the assumption that MsmK contributes to multiple carbohydrates in S. suis.


MsmK, an ATPase, Contributes to Utilization of Multiple Carbohydrates and Host Colonization of Streptococcus suis.

Tan MF, Gao T, Liu WQ, Zhang CY, Yang X, Zhu JW, Teng MY, Li L, Zhou R - PLoS ONE (2015)

Western blot analysis of MsmK expression in SC-19 cultured in different carbohydrates.(A) Strain SC-19 was cultured in CDM supplemented with 1% (g/v) diverse carbohydrates to log phase. The supernatant of cell lysate was disposed for immunoblot analysis with MsmK or Enolase polyclonal antibodies. Enolase was used as an internal reference. (B) The blots were scanned, and the resultant graphics were quantified with ImageJ2x. Fold change is represented as a comparison to cells grown in glucose, and the expression levels of Enolase as internal references. Data are shown as means ± SEM from at least three independent experiments. Statistical significance was tested by a Two-way ANOVA test (ns, P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4519317&req=5

pone.0130792.g005: Western blot analysis of MsmK expression in SC-19 cultured in different carbohydrates.(A) Strain SC-19 was cultured in CDM supplemented with 1% (g/v) diverse carbohydrates to log phase. The supernatant of cell lysate was disposed for immunoblot analysis with MsmK or Enolase polyclonal antibodies. Enolase was used as an internal reference. (B) The blots were scanned, and the resultant graphics were quantified with ImageJ2x. Fold change is represented as a comparison to cells grown in glucose, and the expression levels of Enolase as internal references. Data are shown as means ± SEM from at least three independent experiments. Statistical significance was tested by a Two-way ANOVA test (ns, P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001).
Mentions: Since MsmK is suspected to energize the MsmEFG and MalXCD transporters in S. suis for multiple carbohydrates transport, it was supposed that the levels of MsmK expressed in CDM with diverse carbohydrates aforementioned would be induced in some degree. To obtain the relative expression levels of MsmK, SC-19 was cultured in CDM supplemented with different carbohydrates as sole carbon source. MsmK expression was evaluated by Western blot (Fig 5A). The experimental images (Fig 5A) were quantified by using ImageJ2x software. The amount of MsmK expressed in SC-19 cultured in CDM with glucose was defined as one fold. As expected, expression of MsmK increased on different levels (Fig 5B). According to the result, expression of MsmK in 1% melibiose, glycogen and raffinose were significantly increased (p ≤ 0.01). In contrast, levels of MsmK in 1% maltotriose and maltotraose had fewer changes, but still higher than that in 1% glucose or maltose. MsmK expression induced by these carbohydrates further supports the assumption that MsmK contributes to multiple carbohydrates in S. suis.

Bottom Line: Genetic and biochemistry studies revealed that the MsmK was responsible for the utilization of raffinose, melibiose, maltotetraose, glycogen and maltotriose.In infected mice, the msmK-deletion mutant showed significant defects of survival and colonization when compared with its parental and complementary strains.Taken together, MsmK is an ATPase that contributes to multiple carbohydrates utilization and host colonization of S. suis.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China.

ABSTRACT
Acquisition and metabolism of carbohydrates are essential for host colonization and pathogenesis of bacterial pathogens. Different bacteria can uptake different lines of carbohydrates via ABC transporters, in which ATPase subunits energize the transport though ATP hydrolysis. Some ABC transporters possess their own ATPases, while some share a common ATPase. Here we identified MsmK, an ATPase from Streptococcus suis, an emerging zoonotic bacterium causing dead infections in pigs and humans. Genetic and biochemistry studies revealed that the MsmK was responsible for the utilization of raffinose, melibiose, maltotetraose, glycogen and maltotriose. In infected mice, the msmK-deletion mutant showed significant defects of survival and colonization when compared with its parental and complementary strains. Taken together, MsmK is an ATPase that contributes to multiple carbohydrates utilization and host colonization of S. suis. This study gives new insight into our understanding of the carbohydrates utilization and its relationship to the pathogenesis of this zoonotic pathogen.

No MeSH data available.


Related in: MedlinePlus