Characterization of stm3030 and stm3031 genes of Salmonella enterica serovar Typhimurium in relation to cephalosporin resistance

Autor: Tsuey Ching Yang, Ying Hsiu Lin, Wensi S. Hu
Rok vydání: 2019
Předmět:
0301 basic medicine
Microbiology (medical)
medicine.drug_class
030106 microbiology
Cephalosporin
lcsh:QR1-502
lac operon
Microbial Sensitivity Tests
Serogroup
lcsh:Microbiology
03 medical and health sciences
0302 clinical medicine
Bacterial Proteins
Western blot
Drug Resistance
Multiple
Bacterial

Escherichia coli
medicine
Humans
Immunology and Allergy
030212 general & internal medicine
Promoter Regions
Genetic

Gene
Cephalosporin Resistance
General Immunology and Microbiology
medicine.diagnostic_test
biology
Chemistry
Ceftriaxone
Salmonella enterica
Promoter
Gene Expression Regulation
Bacterial

General Medicine
Chromosomes
Bacterial

biology.organism_classification
Molecular biology
Recombinant Proteins
Infectious Diseases
Genes
Bacterial

Bacterial outer membrane
Gene Deletion
Bacterial Outer Membrane Proteins
Zdroj: Journal of Microbiology, Immunology and Infection, Vol 52, Iss 2, Pp 282-288 (2019)
ISSN: 1684-1182
DOI: 10.1016/j.jmii.2018.09.008
Popis: 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
Databáze: OpenAIRE