Characterization of stm3030 and stm3031 genes of Salmonella enterica serovar Typhimurium in relation to cephalosporin resistance
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(Δstm3031). How stm3030 and stm3031 contributing to antibiotic resist...
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| Format: | Article |
| Language: | English |
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Elsevier
2019-04-01
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| Series: | Journal of Microbiology, Immunology and Infection |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1684118218304134 |
| _version_ | 1819017077622571008 |
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| author | Ying-Hsiu Lin Tsuey-Ching Yang Wensi S. Hu |
| author_facet | Ying-Hsiu Lin Tsuey-Ching Yang Wensi S. Hu |
| author_sort | Ying-Hsiu Lin |
| collection | DOAJ |
| description | 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(Δstm3031). How stm3030 and stm3031 contributing to antibiotic resistance was investigated. Methods: The level of STM3030 protein in R200(Δstm3031) were compared between 01–4, R200, and R200(Δstm3031) by 2-DE analysis. The stm3030 gene deleted strain, R200(Δstm3030), was generated by the one-step inactivation chromosome gene method. The various antibiotic susceptibility of strains 01–4, R200, R200(Δstm3031) and R200(Δstm3030) 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(Δstm3030) 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(Δstm3030) 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. Keywords: Cephalosporin resistance, Salmonella Typhimurium, stm3030, stm3031 |
| first_indexed | 2024-12-21T02:57:48Z |
| format | Article |
| id | doaj.art-24aecc4cf8f94dd4a21364fe0a21d73b |
| institution | Directory Open Access Journal |
| issn | 1684-1182 |
| language | English |
| last_indexed | 2024-12-21T02:57:48Z |
| publishDate | 2019-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Microbiology, Immunology and Infection |
| spelling | doaj.art-24aecc4cf8f94dd4a21364fe0a21d73b2022-12-21T19:18:16ZengElsevierJournal of Microbiology, Immunology and Infection1684-11822019-04-01522282288Characterization of stm3030 and stm3031 genes of Salmonella enterica serovar Typhimurium in relation to cephalosporin resistanceYing-Hsiu Lin0Tsuey-Ching Yang1Wensi S. Hu2Department of Biotechnology and Laboratory Science in Medicine, School of Biomedical Science and Engineering, National Yang-Ming University, Taipei, TaiwanCorresponding author.; Department of Biotechnology and Laboratory Science in Medicine, School of Biomedical Science and Engineering, National Yang-Ming University, Taipei, TaiwanCorresponding author. Department of Biotechnology and Laboratory Science in Medicine, School of Biomedical Science and Engineering, National Yang Ming University, No. 155, Sec. 2, Linong St., Beitou District, Taipei City 112, Taiwan. Fax: +886 2 2826 4092.; Department of Biotechnology and Laboratory Science in Medicine, School of Biomedical Science and Engineering, National Yang-Ming University, Taipei, TaiwanBackground/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(Δstm3031). How stm3030 and stm3031 contributing to antibiotic resistance was investigated. Methods: The level of STM3030 protein in R200(Δstm3031) were compared between 01–4, R200, and R200(Δstm3031) by 2-DE analysis. The stm3030 gene deleted strain, R200(Δstm3030), was generated by the one-step inactivation chromosome gene method. The various antibiotic susceptibility of strains 01–4, R200, R200(Δstm3031) and R200(Δstm3030) 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(Δstm3030) 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(Δstm3030) 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. Keywords: Cephalosporin resistance, Salmonella Typhimurium, stm3030, stm3031http://www.sciencedirect.com/science/article/pii/S1684118218304134 |
| spellingShingle | Ying-Hsiu Lin Tsuey-Ching Yang Wensi S. Hu Characterization of stm3030 and stm3031 genes of Salmonella enterica serovar Typhimurium in relation to cephalosporin resistance Journal of Microbiology, Immunology and Infection |
| title | Characterization of stm3030 and stm3031 genes of Salmonella enterica serovar Typhimurium in relation to cephalosporin resistance |
| title_full | Characterization of stm3030 and stm3031 genes of Salmonella enterica serovar Typhimurium in relation to cephalosporin resistance |
| title_fullStr | Characterization of stm3030 and stm3031 genes of Salmonella enterica serovar Typhimurium in relation to cephalosporin resistance |
| title_full_unstemmed | Characterization of stm3030 and stm3031 genes of Salmonella enterica serovar Typhimurium in relation to cephalosporin resistance |
| title_short | Characterization of stm3030 and stm3031 genes of Salmonella enterica serovar Typhimurium in relation to cephalosporin resistance |
| title_sort | characterization of stm3030 and stm3031 genes of salmonella enterica serovar typhimurium in relation to cephalosporin resistance |
| url | http://www.sciencedirect.com/science/article/pii/S1684118218304134 |
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