Abstract
Antibiotic resistance genes are usually tightly controlled by transcription factors and RNA regulatory elements including sRNAs, riboswitches, and attenuators, and their expression is activated to respond to antibiotic exposure. In previous work, we revealed that the rppA gene is regulated by attenuator LRR and two mistranslation products in Bacillus thuringiensis BMB171. However, its function and promoter regulation is still not precise. In this study, we demonstrated that the encoding product of the rppA gene acts as an ARE1 ABC-F protein and confers resistance to antibiotics virginiamycin M1 and lincomycin when overexpressed. Besides the reported attenuator LRR, the expression of the rppA gene is controlled by the sigma factor SigA and a global transcription factor CcpA. Consequently, its promoter activity is mainly maintained at the stationary phase of cell growth and inhibited in the presence of glucose. Our study revealed the function and regulation of the rppA gene in detail.
Key points
• The RppA protein acts as an ARE1 ABC-F protein
• The rppA gene confers resistance to antibiotics virginiamycin M1 and lincomycin when overexpressed
• The expression of the rppA gene is regulated by the sigma factor SigA and the pleiotropic regulator CcpA
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Acknowledgements
We are very grateful to Dr. Jin He (Huazhong Agricultural University) for providing plasmid pHT1K and pRP1028.
Funding
This work was supported by the National Key Research and Development Program of China (No. 2018YFA0900100), and the National Natural Science Foundation of China (No. 31670081).
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X.C. and J.C. designed the study; X.C. carried out most of the experimental work and analyzed the data; X.L. constructed the plasmids; Q.J. conducted EMSA experiments; Y.Z. and B.Y. conducted in vitro transcription experiments and analyzed some data; J.C. and X.C. wrote the manuscript. All authors read and approved the manuscript.
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Cai, X., Li, X., Qin, J. et al. Gene rppA co-regulated by LRR, SigA, and CcpA mediates antibiotic resistance in Bacillus thuringiensis. Appl Microbiol Biotechnol 106, 5687–5699 (2022). https://doi.org/10.1007/s00253-022-12090-y
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DOI: https://doi.org/10.1007/s00253-022-12090-y