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Peptidoglycan recognition proteins kill bacteria by activating protein-sensing two-component systems

Nature Medicine volume 17, pages 676–683 (2011)Cite this article

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Abstract

Mammalian peptidoglycan recognition proteins (PGRPs), similar to antimicrobial lectins, bind the bacterial cell wall and kill bacteria through an unknown mechanism. We show that PGRPs enter the Gram-positive cell wall at the site of daughter cell separation during cell division. In Bacillus subtilis, PGRPs activate the CssR-CssS two-component system that detects and disposes of misfolded proteins that are usually exported out of bacterial cells. This activation results in membrane depolarization, cessation of intracellular peptidoglycan, protein, RNA and DNA synthesis, and production of hydroxyl radicals, which are responsible for bacterial death. PGRPs also bind the outer membrane of Escherichia coli and activate the functionally homologous CpxA-CpxR two-component system, which kills the bacteria. We exclude other potential bactericidal mechanisms, including inhibition of extracellular peptidoglycan synthesis, hydrolysis of peptidoglycan and membrane permeabilization. Thus, we reveal a previously unknown mechanism by which innate immunity proteins that bind the cell wall or outer membrane exploit the bacterial stress defense response to kill bacteria.

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Figure 1: PGRPs inhibit an intracellular step in peptidoglycan synthesis and localize to the newly formed cell separation site.
Figure 2: PGRPs inhibit protein, RNA and DNA synthesis.
Figure 3: PGRPs induce membrane depolarization, [OH]• formation and HtrA expression through the CssR-CssS two-component system, and [OH]• is responsible for PGRP-induced killing.
Figure 4: PGRPs kill B. subtilis through the CssR-CssS two-component system.
Figure 5: PGRPs kill E. coli through the CpxA-CpxR two-component system and bind the entire outer membrane.
Figure 6: Mammalian PGRPs exploit bacterial defense mechanism to kill bacteria.

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Acknowledgements

We are grateful to J.M. van Dijl, O.P. Kuipers and V.P. Kontinen and their associates J. Zweers and S. Holsappel (University of Groningen and National Institute of Health and Welfare Finland), J. Sekiguchi (Shinshu University) and S.J. Foster (University of Sheffield) for providing B. subtilis mutants; to J.J. Collins and M.A. Kohanski (Boston University) for providing E. coli mutants; to J.M. van Dijl and his associates T. Kouwen and M. Sibbald (University of Groningen) for the pGDL48 plasmid and its sequence; to M. Wang for analyzing samples by mass spectrometry; and to Huvepharma for providing moenomycin. This work was supported by the US Public Health Service grants from the US National Institutes of Health AI073290 and AI028797 to R.D. and D.G. and GM061761 to G.-J.B.

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  1. Des Raj Kashyap and Minhui Wang: These authors contributed equally to this work.

Authors and Affiliations

  1. Indiana University School of Medicine–Northwest, Gary, Indiana, USA

    Des Raj Kashyap, Minhui Wang, Li-Hui Liu, Dipika Gupta & Roman Dziarski

  2. University of Georgia, Athens, Georgia, USA

    Geert-Jan Boons

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Contributions

D.R.K., M.W., D.G. and R.D. designed the experiments, D.R.K., M.W. and R.D. performed the experiments, L.-H.L. and D.R.K. obtained and purified the proteins, G.-J.B. synthesized muramyl peptides, and R.D. wrote the manuscript.

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Correspondence to Roman Dziarski.

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Supplementary Figures 1–8, Supplementary Results, Supplementary Discussion, Supplementary Methods and Supplementary Table 1 (PDF 1852 kb)

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Kashyap, D., Wang, M., Liu, LH. et al. Peptidoglycan recognition proteins kill bacteria by activating protein-sensing two-component systems. Nat Med 17, 676–683 (2011). https://doi.org/10.1038/nm.2357

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