Abstract
The MarR family, as multiple antibiotic resistance regulators, is associated with the resistance of organisms to unfavorable conditions. MarR family extracellular polymeric substances (EPS)–associated transcriptional regulator (EpsRAc) was closely associated with copper resistance in Acidithiobacillus caldus (A. caldus). Transcriptional analysis showed high activity of the epsR promoter (PI) in Escherichia coli and differential response to metal ions. The copper content and UV absorption spectrum of the co-purified protein did not increase, but a stoichiometry of 0.667 mol Cu(I) per EpsRAc monomer was observed in vitro in copper titration experiments, suggesting that Cu(II) acts with low affinity in binding to the EpsRAc protein. Electrophoretic mobility shift assays (EMSA) demonstrated that EpsRAc could bind to its own promoter in vitro, and the binding region was the palindrome sequence TGTTCATCGTGTGTGAGCACACA. EpsRAc negatively regulated its own gene expression, whereas Cu(II) mitigates this negative effect. EpsRAc did not bind to other neighboring gene promoters. Finally, we developed a working model to illustrate the regulatory mechanism of A. caldus in response to extreme copper stress.
Key points
• Identification of a MarR family EPS-associated transcriptional regulator, named EpsRAc.
• Cu(I) can bind to the EpsRAc protein with low affinity.
• EpsRAc negatively regulates the expression of epsR, and Cu(II) can alleviate this negative regulation.
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Funding
This study was supported by grants from the National Natural Science Foundation of China (Nos. 21878128; 21776113; 21606110; 31701582), the funding of Key Laboratory of Industrial Biotechnology, Ministry of Education (KLIBKF202005), the funding of Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education (KLCCB-KF202001), the Fundamental Research Funds for the Central Universities (No. 2050205), the Priority Academic Program Development of Jiangsu Higher Education Institutions, and Program of Introducing Talents of Discipline to Universities (No. 111–2-06).
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YY and SF designed the research and analyzed the data. YY performed the experiments and wrote the first draft of the manuscript. SF, HY, and YT contributed to manuscript revision, read, and approved the submitted version. All authors read and approved the manuscript.
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Yin, Y., Tong, Y., Yang, H. et al. EpsRAc is a copper-sensing MarR family transcriptional repressor from Acidithiobacillus caldus. Appl Microbiol Biotechnol 106, 3679–3689 (2022). https://doi.org/10.1007/s00253-022-11971-6
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DOI: https://doi.org/10.1007/s00253-022-11971-6