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PMID:22985829
| Citation |
Filipic, B, Golic, N, Jovcic, B, Tolinacki, M, Bay, DC, Turner, RJ, Antic-Stankovic, J, Kojic, M and Topisirovic, L (2013) The cmbT gene encodes a novel major facilitator multidrug resistance transporter in Lactococcus lactis. Res. Microbiol. 164:46-54 |
|---|---|
| Abstract |
Functional characterization of the multidrug resistance CmbT transporter was performed in Lactococcus lactis. The cmbT gene is predicted to encode an efflux protein homologous to the multidrug resistance major facilitator superfamily. The cmbT gene (1377 bp) was cloned and overexpressed in L. lactis NZ9000. Results from cell growth studies revealed that the CmbT protein has an effect on host cell resistance to lincomycin, cholate, sulbactam, ethidium bromide, Hoechst 33342, sulfadiazine, streptomycin, rifampicin, puromycin and sulfametoxazole. Moreover, in vivo transport assays showed that overexpressed CmbT-mediated extrusion of ethidium bromide and Hoechst 33342 was higher than in the control L. lactis NZ9000 strain. CmbT-mediated extrusion of Hoechst 33342 was inhibited by the ionophores nigericin and valinomycin known to dissipate proton motive force. This indicates that CmbT-mediated extrusion is based on a drug-proton antiport mechanism. Taking together results obtained in this study, it can be concluded that CmbT is a novel major facilitator multidrug resistance transporter candidate in L. lactis, with a possible signaling role in sulfur metabolism. |
| Links |
PubMed Online version:10.1016/j.resmic.2012.09.003 |
| Keywords |
Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Cloning, Molecular; Drug Resistance, Multiple, Bacterial/genetics; Gene Expression; Lactococcus lactis/genetics; Lactococcus lactis/metabolism; Membrane Transport Proteins/genetics; Membrane Transport Proteins/metabolism; Microbial Sensitivity Tests; Recombinant Proteins/genetics; Recombinant Proteins/metabolism; Sulfur/metabolism |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
|
enables |
GO:0015299: solute:proton antiporter activity |
ECO:0000314: direct assay evidence used in manual assertion |
F |
Seeded From UniProt |
complete | |||
| GO:0015299: solute:proton antiporter activity |
ECO:0000314: direct assay evidence used in manual assertion |
F |
Table 2 shows that expression of the cmbT gene in the CT501 strain confers resistance to a range of antibiotics. Figure 3A shows the decreased intercalation of ethidium bromide into DNA (less fluorescence) in the CT501 strain. Figure 3B shows that this does not change in the presence of an ATPase inhibitor, implying the transport of ethidium bromide out of the cell is proton motive force driven. |
complete | ||||
Notes
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