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Pseudomonas aeruginosa regulator PvrA binds simultaneously to multiple pseudo-palindromic sites for efficient transcription activation

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Abstract

Tetracycline repressor (TetR) family regulators (TFRs) are the largest group of DNA-binding transcription factors and are widely distributed in bacteria and archaea. TFRs play vital roles in controlling the expression of various genes and regulating diverse physiological processes. Recently, a TFR protein Pseudomonas virulence regulator A (PvrA), was identified from Pseudomonas aeruginosa as the transcriptional activator of genes involved in fatty acid utilization and bacterial virulence. Here, we show that PvrA can simultaneously bind to multiple pseudo-palindromic sites and upregulate the expression levels of target genes. Cryo-electron microscopy (cryo-EM) analysis indicates the simultaneous DNA recognition mechanism of PvrA and suggests that the bound DNA fragments consist of a distorted B-DNA double helix. The crystal structure and functional analysis of PvrA reveal a hinge region that secures the correct domain motion for recognition of the promiscuous promoter. Additionally, our results showed that mutations disrupting the regulatory hinge region have differential effects on biofilm formation and pyocyanin biosynthesis, resulting in attenuated bacterial virulence. Collectively, these findings will improve the understanding of the relationship between the structure and function of the TetR family and provide new insights into the mechanism of regulation of P. aeruginosa virulence.

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Acknowledgement

This work was supported by the Ministry of Science and Technology of China (2022YFC2303700, 2021YFA1301900). The National Natural Science Foundation of China (81871615, 32222040, 32070049), Tianjin Synthetic Biotechnology Innovation Capacity Improvement Action (TSBICIP-KJGG-008). And this work was supported in part by a Tibet Science Foundation grant (XZ202001ZY0036 N) to Yonghong Zhou. Cryo-EM data were collected at SKLB West China Cryo-EM Center and processed at SKLB Duyu High Performance Computing Center in Sichuan University. We thank National Center for Protein Sciences Shanghai (NCPSS) beamlines BL18U and BL19U allowance. We thank the staff at BL17B1/BL18U1/BL19U1 beamlines at SSRF of the National Facility for Protein Science in Shanghai (NFPS), Shanghai Advanced Research Institute, Chinese Academy of Sciences, for providing technical support in X-ray diffraction data collection and analysis.

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Authors and Affiliations

  1. Center of Infectious Diseases, Division of Infectious Diseases in State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, 610041, China

    Yibo Zhu, Bingnan Luo, Xingyu Mou, Yongbo Luo, Youfu Luo, Hong Tang, Zhaoming Su & Rui Bao

  2. Department of Geriatrics and National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University, Chengdu, 610041, China

    Zhaoming Su

  3. Key Laboratory of Biodiversity and Environment on the Qinghai-Tibetan Plateau, Ministry of Education, School of Ecology and Environment, Tibet University, Lhasa, 850000, China

    Yonghong Zhou

  4. Shanghai Advanced Research Institute, Chinese Academy of Sciences, Shanghai, 201210, China

    Bo Sun

  5. College of Life Science, Sichuan Normal University, Chengdu, 610101, China

    Yingjie Song

  6. Accurate Biotechnology (Hunan) Co., Ltd, Changsha, 410006, China

    Yibo Zhu

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  1. Yibo Zhu
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Pseudomonas aeruginosa regulator PvrA binds simultaneously to multiple pseudo-palindromic sites for efficient transcription activation

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Zhu, Y., Luo, B., Mou, X. et al. Pseudomonas aeruginosa regulator PvrA binds simultaneously to multiple pseudo-palindromic sites for efficient transcription activation. Sci. China Life Sci. 67, 900–912 (2024). https://doi.org/10.1007/s11427-022-2363-y

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