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CusS-CusR Two-Component System Mediates Tigecycline Resistance in Carbapenem-Resistant . | LitMetric
Background : The increase in carbapenem-resistant (CRKP), especially the emergence of tigecycline-resistant (KP), is a serious public health concern. However, the underlying mechanism of tigecycline resistance is unclear. In this study, we evaluated the role of the CusS-CusR two-component system (TCS), which is associated with copper/silver resistance, in tigecycline resistance in CRKP.Methods : Following the evolution of tigecycline-resistant KP, the minimum inhibitory concentrations of tigecycline were determined using the micro-broth dilution method. RNA sequencing and data analysis were performed to identify differentially expressed genes. Quantitative PCR (qPCR) was performed to verify the genes of interest. Genes associated with tigecycline resistance, such as , , and , were detected by PCR, and then mutants were confirmed by sequencing. Additionally, the efflux pump-associated genes , , , , and were also analyzed by qPCR. was deleted and complemented by the suicide vector pKO3-Km plasmid and pGEM-T-easy plasmid, respectively.Results : Nine strains of KP were evaluated in our study. Strains and were evolved from , , and were evolved from , and and were evolved from . The tigecycline minimum inhibitory concentration for , , and C1 was 0.5 μg/mL; that for , , and was 16.0 μg/mL; and that for , , and was 32.0 μg/mL. RNA-sequencing and qPCR confirmed that the differentially expressed genes , , , , , , and showed higher expression in and than in . Genes related to the efflux pump AcrAB-TolC showed higher expression in and than in . No mutants of , , or were detected. , , , , and did not show increased expression in any group. After deletion and complementation of cusR among C3, the MIC of tigecycline decreased to 4 μg/mL, and then recovered to 32 μg/mL. The expression of , correspondingly decreased and increased significantly.Conclusion : In addition to its primary function in resistance to copper/silver, the CusS-CusR two-component system is associated with CRKP resistance to tigecycline.
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