Abstract
Objectives
To identify proteins that may be associated with antibiotic resistance in the multidrug-resistant Salmonella enterica D14, by constructing proteomic profiles using mass spectrometry-based label-free quantitative proteomics (LFQP).
Results
D14 was cultured with four antibiotics (ampicillin, nalidixic acid, streptomycin, and tetracycline) separately. Subsequently, the findings from an equal combination of the four cultures were compared with the profile of sensitive S. enterica 104. 2255 proteins, including 149 differentially up-regulated proteins, were identified. Many of these up-regulated proteins were associated with flagellar assembly and chemotaxis, two-component system, amino acid metabolism, β-lactam resistance, and transmembrane transport. A subset of 10 genes was evaluated via quantitative real-time PCR (qPCR), followed by the construction of cheR, fliS, fliA, arnA, and yggT deletion mutants. Only the yggT-deleted D14 mutant showed decrease in streptomycin resistance, whereas the other deletions had no effect. Furthermore, complementation of yggT and the overexpression of yggT in S. enterica ATCC 14028 increased the streptomycin resistance. Additionally, spot dilution assay results confirmed that Salmonella strains, harboring yggT, exhibited an advantage in the presence of streptomycin.
Conclusions
The above proteomic and mutagenic analyses revealed that yggT is involved in streptomycin resistance in S. enterica.
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Acknowledgements
We acknowledge Prof. Yahong Liu (South China Agricultural University) for providing Salmonella strains D14 and 104. We acknowledge Pro. Sheng Yang (Chinese Academy of Sciences) for providing the plasmid pREDTKI. We acknowledge Prof. Roy Curtiss, III (The University of Florida) for providing the plasmid pRE112.
Supplementary Information
Supplementary Table 1—Plasmids and primers used in this work.
Supplementary Table 2—Identification of differentially up-regulated proteins (n = 149).
Supplementary Table 3—Identification of differentially down-regulated proteins (n = 26).
Funding
This work was funded by grants from the National Natural Science Foundation of China (No. 31802246), and Science and Technology Program of Guangzhou (No. 201804010207).
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Liang, Y., Deng, F., Mu, P. et al. Quantitative proteomics implicates YggT in streptomycin resistance in Salmonella enterica serovar Enteritidis. Biotechnol Lett 43, 919–932 (2021). https://doi.org/10.1007/s10529-021-03083-4
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DOI: https://doi.org/10.1007/s10529-021-03083-4