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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

MsmK is an ATPase.(A) Genetic map of the loci encoding predicated carbohydrate ATPase, MsmK, in SC84. Large arrows represent open reading frames and their direction of transcription. Small arrow indicates the promoter of the gene msmK and its transcription direction. (B) SDS-PAGE of purified recombinant protein MsmK. Before electrophoresis, the proteins were exposed to no DTT (left lane,-) or 2 mM DTT (right lane, +DTT). Purified MsmK was formed by a certain amount of monomers (42 KDa) and dimmers (~80 KDa). (C) ATPase activity of MsmK. Protein was incubated with 1 mM ATP for 60 min, and the generation of ADP was quantified. The mixture with BSA protein was used as a negative control. Data represent the average ± SD of at least three independent repeats.
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pone.0130792.g002: MsmK is an ATPase.(A) Genetic map of the loci encoding predicated carbohydrate ATPase, MsmK, in SC84. Large arrows represent open reading frames and their direction of transcription. Small arrow indicates the promoter of the gene msmK and its transcription direction. (B) SDS-PAGE of purified recombinant protein MsmK. Before electrophoresis, the proteins were exposed to no DTT (left lane,-) or 2 mM DTT (right lane, +DTT). Purified MsmK was formed by a certain amount of monomers (42 KDa) and dimmers (~80 KDa). (C) ATPase activity of MsmK. Protein was incubated with 1 mM ATP for 60 min, and the generation of ADP was quantified. The mixture with BSA protein was used as a negative control. Data represent the average ± SD of at least three independent repeats.

Mentions: Like most operons encoding CUT1 transporters in Streptococci species (such as S. pneumoniae in Fig 1C and 1F), there is no ATPase gene in the operon. Given that ATPase is an essential component of an ABC transporter to energize the substrate transport, an ATPase gene must exist in other locus or loci in the genome of these Streptococci species. It is known that in S. mutans, MsmK and MalK function as ATPase for the MsmEFG and MalXFG transporters, respectively [35], while MsmK energizes multiple carbohydrates transporters in S. pneumoniae including the MalXCD transporter [32]. To identify the ATPase which energizes the MsmEFG and MalXCD transporters in S. suis, we screened S. suis genome sequences available in GenBank and found that the protein encoded by SSUSC84_1724 in the genome of S. suis strain SC84 showed the highest similarity to the MsmK of S. pneumoniae (79.5%) and S. mutans (78.5%) (Fig 2A) [30,32,33,35]. It contains the typical ATPase domains as MsmK: Walker A, Q-loop, ABC transporter signature motif, Walker B, D-loop and H-loop (S1 Fig) [27,52]. Taken together, we speculate that the SSUSC84_1724 gene encodes an ATPase for the MsmEFG and MalXCD transporters of S. suis, and rename it as MsmK.


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)

MsmK is an ATPase.(A) Genetic map of the loci encoding predicated carbohydrate ATPase, MsmK, in SC84. Large arrows represent open reading frames and their direction of transcription. Small arrow indicates the promoter of the gene msmK and its transcription direction. (B) SDS-PAGE of purified recombinant protein MsmK. Before electrophoresis, the proteins were exposed to no DTT (left lane,-) or 2 mM DTT (right lane, +DTT). Purified MsmK was formed by a certain amount of monomers (42 KDa) and dimmers (~80 KDa). (C) ATPase activity of MsmK. Protein was incubated with 1 mM ATP for 60 min, and the generation of ADP was quantified. The mixture with BSA protein was used as a negative control. Data represent the average ± SD of at least three independent repeats.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0130792.g002: MsmK is an ATPase.(A) Genetic map of the loci encoding predicated carbohydrate ATPase, MsmK, in SC84. Large arrows represent open reading frames and their direction of transcription. Small arrow indicates the promoter of the gene msmK and its transcription direction. (B) SDS-PAGE of purified recombinant protein MsmK. Before electrophoresis, the proteins were exposed to no DTT (left lane,-) or 2 mM DTT (right lane, +DTT). Purified MsmK was formed by a certain amount of monomers (42 KDa) and dimmers (~80 KDa). (C) ATPase activity of MsmK. Protein was incubated with 1 mM ATP for 60 min, and the generation of ADP was quantified. The mixture with BSA protein was used as a negative control. Data represent the average ± SD of at least three independent repeats.
Mentions: Like most operons encoding CUT1 transporters in Streptococci species (such as S. pneumoniae in Fig 1C and 1F), there is no ATPase gene in the operon. Given that ATPase is an essential component of an ABC transporter to energize the substrate transport, an ATPase gene must exist in other locus or loci in the genome of these Streptococci species. It is known that in S. mutans, MsmK and MalK function as ATPase for the MsmEFG and MalXFG transporters, respectively [35], while MsmK energizes multiple carbohydrates transporters in S. pneumoniae including the MalXCD transporter [32]. To identify the ATPase which energizes the MsmEFG and MalXCD transporters in S. suis, we screened S. suis genome sequences available in GenBank and found that the protein encoded by SSUSC84_1724 in the genome of S. suis strain SC84 showed the highest similarity to the MsmK of S. pneumoniae (79.5%) and S. mutans (78.5%) (Fig 2A) [30,32,33,35]. It contains the typical ATPase domains as MsmK: Walker A, Q-loop, ABC transporter signature motif, Walker B, D-loop and H-loop (S1 Fig) [27,52]. Taken together, we speculate that the SSUSC84_1724 gene encodes an ATPase for the MsmEFG and MalXCD transporters of S. suis, and rename it as MsmK.

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