[No authors listed]
BACKGROUND/PURPOSE:The outer membrane protein STM3031 had been shown to confer Salmonella enterica serovar Typhimurium resistance to ceftriaxone. In this study, the STM3030 was increased in strain R200 and decreased in strain R200(Îstm3031). How stm3030 and stm3031 contributing to antibiotic resistance was investigated. METHODS:The level of STM3030 protein in R200(Îstm3031) were compared between 01-4, R200, and R200(Îstm3031) by 2-DE analysis. The stm3030 gene deleted strain, R200(Îstm3030), was generated by the one-step inactivation chromosome gene method. The various antibiotic susceptibility of strains 01-4, R200, R200(Îstm3031) and R200(Îstm3030) were determined by agar dilutions assays and E-test. The co-transcription of stm3031 and stm3030 were determined by RT-PCR. The promoter activities of these two genes fused with LacZ were determined. The binding of the regulatory protein BaeR on the promoter of both genes was detected by EMSA. The interaction between STM3030 and STM3031 proteins was determined by GST pull-down assay. RESULTS:Strain R200(Îstm3030) displayed a 32- to 64-fold reduction in resistance to cephalosporin drugs. Transcription analyses revealed that stm3030 and stm3031 are independent genes and that the promoter of stm3030 is stronger than that of stm3031. The regulator BaeR binds to the promoter region of stm3031 but not that of stm3030. The STM3031 decreased in R200(Îstm3030) compared to R200 by western blot analysis. The pull-down assay revealed that STM3030 and STM3031 bind to each other. CONCLUSION:Our data indicate that STM3030 has a chaperone-like activity and may modulate or stabilize STM3031, leading to resistance of S. enterica serovar Typhimurium to cephalosporin drugs.
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