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Comparative Study
. 2019 Jul 2;15(1):224.
doi: 10.1186/s12917-019-1959-9.

Comparative proteomic analysis reveals drug resistance of Staphylococcus xylosus ATCC700404 under tylosin stress

Xin Liu  1 , Jinpeng Wang  1 , Mo Chen  1 , Ruixiang Che  1 , Wenya Ding  1 , Fei Yu  1 , Yonghui Zhou  1 , Wenqiang Cui  1 , Xing Xiaoxu  1 , Bello-Onaghise God'spower  1 , Yanhua Li  2
Affiliations
Comparative Study

Comparative proteomic analysis reveals drug resistance of Staphylococcus xylosus ATCC700404 under tylosin stress

Xin Liu et al. BMC Vet Res. .

Abstract

Background: As a kind of opportunist pathogen, Staphylococcus xylosus (S. xylosus) can cause mastitis. Antibiotics are widely used for treating infected animals and tylosin is a member of such group. Thus, the continuous use of antibiotics in dairy livestock enterprise will go a long way in increasing tylosin resistance. However, the mechanism of tylosin-resistant S. xylosus is not clear. Here, isobaric tag for relative and absolute quantitation (iTRAQ)-based quantitative proteomics methods was used to find resistance-related proteins.

Results: We compared the differential expression of S. xylosus in response to tylosin stress by iTRAQ. A total of 155 proteins (59 up-regulated, 96 down-regulated) with the fold-change of >1.2 or <0.8 (p value ≤0.05) were observed between the S. xylosus treated with 1/2 MIC (0.25 μg/mL) tylosin and the untreated S. xylosus. Bioinformatic analysis revealed that these proteins play important roles in stress-response and transcription. Then, in order to verify the relationship between the above changed proteins and mechanism of tylosin-resistant S. xylosus, we induced the tylosin-resistant S. xylosus, and performed quantitative PCR analysis to verify the changes in the transcription proteins and the stress-response proteins in tylosin-resistant S. xylosus at the mRNA level. The data displayed that ribosomal protein L23 (rplw), thioredoxin(trxA) and Aldehyde dehydrogenase A(aldA-1) are up-regulated in the tylosin-resistant S. xylosus, compared with the tylosin-sensitive strains.

Conclusion: Our findings demonstrate the important of stress-response and transcription in the tylosin resistance of S. xylosus and provide an insight into the prevention of this resistance, which would aid in finding new medicines .

Keywords: Aldehyde dehydrogenase a;; Drug-resistance; Ribosomal protein L23; Staphylococcus xylosus; Thioredoxin; Tylosin; iTRAQ.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Significantly differential proteins of Staphylococcus xylosus ATCC700404 in 0.25 μg/mL tylosin stress using iTRAQ. a The number of altered proteins. b K-means clustering representation of total 155 DEP profiles. The magnitude of the percentage is represented by a color scale (top right) going from low (green) to high (red)
Fig. 2
Fig. 2
Go annotation and KEGG pathway of DEPs: Geneontology terms for subcellular laocation distribution. a cellular component (b) biological process (c) molecular function (d) main KEGG pathway
Fig. 3
Fig. 3
String network of significantly differential proteins of Staphylococcus xylosus ATCC700404 in 0.25 μg/mL tylosin stress. Colored lines between the proteins indicate the various types of interaction evidence. Structure which is drawn in the protein nodes indicated the availability of 3D protein structure information
Fig. 4
Fig. 4
The string network and qPCR analysis of possible tylosin-resistant proteins. a The string network of primary tylosin-resistant proteins with altered expression. b The mRNA levels of two translation and four stress-response related genes were respectively analyzed by qPCR method in tylosin stress and tylosin-senstive Staphylococcus xylosus ATCC700404 and tylosin-resistant Staphylococcus xylosus ATCC700404. T: tylsoin, S.X.: tylosin-senstive Staphylococcus xylosus ATCC700404, S.X.-R: tylosin-resistant Staphylococcus xylosus ATCC700404
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