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ClpXP affects the cell metabolism of Salmonella typhimurium partially in an RpoS-dependent manner

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Abstract

Introduction

ClpXP protease is an important proteolytic system in Salmonella enterica serovar typhimurium (S. typhimurium). Inactivation of ClpXP by deletion of clpP resulted in overproduction of RpoS and a growth defect phenotype. Only one report has indicated that deleting rpoS can restore the growth of a S. typhimurium clpP mutant to the wild-type level. Whether overproduction of RpoS is responsible for the growth deficiency resulting from clpP disruption and how ClpXP affects the cell metabolism of S. typhimurium remain to be elucidated.

Objectives

The aim of this study is to investigate the effect of ClpXP on cell metabolism of S. typhimurium and explore the possible co-effect of RpoS associated with ClpXP in cell metabolism.

Method

We constructed a clpP rpoS double deletion mutant TT-19 (ΔclpP ΔrpoS TT-1) using a two-step phage transduction technique. We then compared the metabolite fingerprints of Salmonella rpoS deletion mutant TT-14 (ΔrpoS TT-1), clpP deletion mutant TT-16 (ΔclpP TT-1), and clpP rpoS double deletion mutant TT-19 (ΔclpP ΔrpoS TT-1) with those of the wild-type strain TT-1 by using gas chromatography coupled with mass spectrometry (GC–MS).

Results

Deletion of rpoS recovered only a part of the growth of Salmonella clpP mutant. Further metabolome analysis indicated that clpP disruption changed the levels of 16 extra- and 19 intracellular substances, while the extracellular concentrations of 4 compounds (serine, l-5-oxoproline, l-glutamic acid, and l-tryptophan) and intracellular concentrations of 10 compounds (l-isoleucine, glycine, serine, l-methionine, l-phenylalanine, malic acid, citric acid, urea, putrescine, and 6-hydroxypurine) returned to their wild-type levels when rpoS was also deleted.

Conclusion

ClpXP affects the cell metabolism of S. typhimurium partially in an RpoS-dependent manner.

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Acknowledgements

This study is supported by the China Postdoctoral Science Foundation (No. 2016M602691), Sichuan Science & Technology Department Foundation (No. 2017JY0241), Young Scholar Research Grants of Sichuan University (No. 2016SCU11005) and Natural Science Foundation of China (No. 31570924).

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Author notes

  1. Tian Tang and Qun Gao have contributed equally to this work and should be listed as first authors.

Authors and Affiliations

  1. West China School of Public Health, Sichuan University, 16#, Section 3, Ren Min Nan Road, Chengdu, 610041, Sichuan, People’s Republic of China

    Tian Tang, Jingyuan Xiong, Xiaofang Pei, Bo Zheng, Xiaoli Zou & Chuan Wang

  2. Avian Diseases Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, 611130, Sichuan, People’s Republic of China

    Qun Gao

  3. Sichuan Entry-Exit Inspection and Quarantine Bureau, Chengdu, 610041, Sichuan, People’s Republic of China

    Hua Lin

  4. Unité des Infections Bactériennes Invasives, Institut Pasteur, Paris, France

    Francis Biville

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  1. Tian Tang
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Contributions

TT, QG, FB, XP and CW designed the experiments. BZ, XZ and JX guided the experiments. TT performed the experiments. TT analyzed the data. TT and FB wrote this paper.

Corresponding author

Correspondence to Chuan Wang.

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Tang, T., Gao, Q., Lin, H. et al. ClpXP affects the cell metabolism of Salmonella typhimurium partially in an RpoS-dependent manner. Metabolomics 13, 157 (2017). https://doi.org/10.1007/s11306-017-1296-6

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