A critical role for peptidoglycan N-deacetylation in Listeria evasion from the host innate immune system - PubMed (original) (raw)
. 2007 Jan 16;104(3):997-1002.
doi: 10.1073/pnas.0609672104. Epub 2007 Jan 10.
Olivier Dussurget, Didier Cabanes, Marie-Anne Nahori, Sandra Sousa, Marc Lecuit, Emmanuel Psylinakis, Vassilis Bouriotis, Jean-Pierre Hugot, Marco Giovannini, Anthony Coyle, John Bertin, Abdelkader Namane, Jean-Claude Rousselle, Nadège Cayet, Marie-Christine Prévost, Viviane Balloy, Michel Chignard, Dana J Philpott, Pascale Cossart, Stephen E Girardin
Affiliations
- PMID: 17215377
- PMCID: PMC1766339
- DOI: 10.1073/pnas.0609672104
A critical role for peptidoglycan N-deacetylation in Listeria evasion from the host innate immune system
Ivo G Boneca et al. Proc Natl Acad Sci U S A. 2007.
Abstract
Listeria monocytogenes is a human intracellular pathogen that is able to survive in the gastrointestinal environment and replicate in macrophages, thus bypassing the early innate immune defenses. Peptidoglycan (PG) is an essential component of the bacterial cell wall readily exposed to the host and, thus, an important target for the innate immune system. Characterization of the PG from L. monocytogenes demonstrated deacetylation of N-acetylglucosamine residues. We identified a PG N-deacetylase gene, pgdA, in L. monocytogenes genome sequence. Inactivation of pgdA revealed the key role of this PG modification in bacterial virulence because the mutant was extremely sensitive to the bacteriolytic activity of lysozyme, and growth was severely impaired after oral and i.v. inoculations. Within macrophage vacuoles, the mutant was rapidly destroyed and induced a massive IFN-beta response in a TLR2 and Nod1-dependent manner. Together, these results reveal that PG N-deacetylation is a highly efficient mechanism used by Listeria to evade innate host defenses. The presence of deacetylase genes in other pathogenic bacteria indicates that PG N-deacetylation could be a general mechanism used by bacteria to evade the host innate immune system.
Conflict of interest statement
The authors declare no conflict of interest.
Figures
Fig. 1.
Characterization of Listeria PG. (A) Each muropeptide peak highlighted by a number was purified by HPLC, desalted, and analyzed by MALDI-TOF. Structural assignment was done by muropeptide fragmentation using MALDI-postsource decay (PSD). The structure of major muropeptides is indicated by full arrows. Structures or substructures in red and black indicate fully acetylated and N-deacetylated moieties, respectively, of the different muropeptides. Peaks 1–22 correspond to monomeric muropeptides, peaks 23- 44 correspond to dimeric muropeptides, and 46 and over correspond to trimeric muropeptides. Approximately 50% of the muropeptides present a glucosamine residue instead of the canonical _N_-acetylglucosamine residue. (B) HPLC analysis of the muropeptide composition of Listeria WT EGDe strain and its pgdA isogenic mutant. Each muropeptide peak was purified by HPLC and analyzed by MALDI-PSD mass spectrometry. Muropeptide peaks indicated with an asterisk correspond to N-deacetylated muropeptides. N-deacetylated muropeptides characteristic of the parental strain EGDe were completely absent from the elution pattern of the pgdA mutant. Glc_N_Ac, N_-acetylglucosamine; Glc_N, glucosamine; M, _N_-acetylmuramic acid, TriPDAP,
l
-alanyl-γ-
d
-glutamyl-_meso_diaminopimelic acid; TriPDAPNH2,
l
-alanyl-γ-
d
-glutamyl-amidated _meso_diaminopimelic acid; TetraPDAP,
l
-alanyl-γ-
d
-glutamyl-_meso_diaminopimelyl-
d
-alanine; TetraPDAPNH2,
l
-alanyl-γ-
d
-glutamyl-amidated _meso_diaminopimelyl-
d
-alanine.
Fig. 2.
Effect of lysozyme on growth and impaired survival in macrophages of the pgdA mutant. Strain EGDe and its isogenic pgdA mutant were grown in BHI media and incubated with lysozyme (10 μg/ml) or the human serum amidase (1 μg/ml). (A) The pgdA mutant was selectively sensitive to the action of lysozyme upon entry into stationary phase. (B) Decrease of the optical density of the pgdA mutant correlated with a loss of viability, whereas the parental strain was insensitive to lysozyme. Lysozyme induced cell rounding of the pgdA mutant (see
SI Fig. 7
). (C and D) RAW264.7 macrophages (C) and PEM (D) were infected with WT EGDe and its pgdA mutant. Sensitivity of the pgdA mutant to lysozyme correlated with its impaired survival in macrophages.
Fig. 3.
Impaired survival of the pgdA mutant in macrophages. (A–D) RAW264.7 cells after 8 h of infection with the parental strain EGDe (A) and the pgdA mutant (B). Impaired survival was correlated with delay in escape of the pgdA mutant (C) from phagosomes compared with the WT strain EGDe (D). (E and F) PEM after 7 h of infection with the parental strain EGDe (E) and the pgdA mutant (F). [Scale bars: 2 μm and 1 μm (Insets).] Impaired survival correlated with delay in escape from phagosomes and bacterial lysis of the pgdA mutant compared with the WT strain EGDe.
Fig. 4.
Impaired virulence of the pgdA mutant in vivo. (A and B) BALB/c and C57/BL6J mice were challenged by i.v. injection with the parental strain EGDe and its pgdA mutant with a sublethal dose (5 × 103 CFU per mouse). After 72 h, mice were killed, and bacterial counts in the liver (A) and the spleen (B) were determined. (C–G) Transgenic human E-cadherin mice were used as model for the oral route of infection, and colonization of several organs was followed after 3, 24, 48, and 72 h after challenge. Interestingly, the pgdA mutant was particularly vulnerable to persistence in the intestinal lumen as assayed by bacterial counts per grams of feces (C). Survival was more robust in the intestine (D) and the mesenteric lymph nodes (E) where colonization was particularly impaired after 72 h. The mutant was impaired in the survival of the liver (F) and spleen (G) at all time points as in the IV model shown in A and B.
Fig. 5.
Enhanced inflammatory response of the pgdA mutant. Cytokine production such as IL-6 (A) and IFN-β (B) by RAW264.7 macrophages was enhanced by the pgdA mutant compared with the parental strain EGDe (see also
SI Fig. 9
). PEM of WT C57/BL6J, Nod1−/−, Nod2−/− TLR2−/− and MyD88−/− mice were infected with strains EGDe and its isogenic pgdA mutant. After 7 h of infection, the inflammatory response was enhanced as measured by the amount of IL-6 (C) and, particularly, IFN-β (D) production. The cytokine response depended mainly on TLR2 and Myd88, although their contribution varied according to the cytokine measured. IFN-β production depended almost entirely on TLR2 in a MyD88-dependent and -independent manner (D). IL-6 production depended only partially on TLR2 and Myd88 (C) and also required Nod1. Surprisingly, Nod1 seemed to function as an inhibitor of IFN-β production, because the pgdA mutant induced 3-fold more IFN-β in Nod1−/− compared with C57/BL6J peritoneal macrophages (D). Note that the role of Nod1 is entirely restricted to the response to the pgdA mutant and does not participate in the response to the WT EGDe strain.
Comment in
- Deception point: peptidoglycan modification as a means of immune evasion.
Bishop JL, Boyle EC, Finlay BB. Bishop JL, et al. Proc Natl Acad Sci U S A. 2007 Jan 16;104(3):691-2. doi: 10.1073/pnas.0611133104. Epub 2007 Jan 12. Proc Natl Acad Sci U S A. 2007. PMID: 17220274 Free PMC article. No abstract available.
Similar articles
- Mutations of the Listeria monocytogenes peptidoglycan N-deacetylase and O-acetylase result in enhanced lysozyme sensitivity, bacteriolysis, and hyperinduction of innate immune pathways.
Rae CS, Geissler A, Adamson PC, Portnoy DA. Rae CS, et al. Infect Immun. 2011 Sep;79(9):3596-606. doi: 10.1128/IAI.00077-11. Epub 2011 Jul 18. Infect Immun. 2011. PMID: 21768286 Free PMC article. - Autolysin amidase of Listeria monocytogenes promotes efficient colonization of mouse hepatocytes and enhances host immune response.
Asano K, Sashinami H, Osanai A, Asano Y, Nakane A. Asano K, et al. Int J Med Microbiol. 2011 Aug;301(6):480-7. doi: 10.1016/j.ijmm.2011.01.002. Epub 2011 Mar 8. Int J Med Microbiol. 2011. PMID: 21388880 - Helicobacter pylori peptidoglycan modifications confer lysozyme resistance and contribute to survival in the host.
Wang G, Lo LF, Forsberg LS, Maier RJ. Wang G, et al. mBio. 2012 Dec 4;3(6):e00409-12. doi: 10.1128/mBio.00409-12. mBio. 2012. PMID: 23221800 Free PMC article. - Listeria monocytogenes infection in the face of innate immunity.
Corr SC, O'Neill LA. Corr SC, et al. Cell Microbiol. 2009 May;11(5):703-9. doi: 10.1111/j.1462-5822.2009.01294.x. Epub 2009 Feb 2. Cell Microbiol. 2009. PMID: 19191786 Review. - Peptidoglycan recognition in innate immunity.
Dziarski R, Gupta D. Dziarski R, et al. J Endotoxin Res. 2005;11(5):304-10. doi: 10.1179/096805105X67256. J Endotoxin Res. 2005. PMID: 16263004 Review.
Cited by
- The hidden base of the iceberg: gut peptidoglycome dynamics is foundational to its influence on the host.
Wheeler R, Gomperts Boneca I. Wheeler R, et al. Gut Microbes. 2024 Jan-Dec;16(1):2395099. doi: 10.1080/19490976.2024.2395099. Epub 2024 Sep 6. Gut Microbes. 2024. PMID: 39239828 Free PMC article. Review. - Molecular Mechanisms of Bacterial Resistance to Antimicrobial Peptides in the Modern Era: An Updated Review.
Tajer L, Paillart JC, Dib H, Sabatier JM, Fajloun Z, Abi Khattar Z. Tajer L, et al. Microorganisms. 2024 Jun 21;12(7):1259. doi: 10.3390/microorganisms12071259. Microorganisms. 2024. PMID: 39065030 Free PMC article. Review. - Mode of action of Akkermansia muciniphila in the intestinal dialogue: role of extracellular proteins, metabolites and cell envelope components.
Segers A, de Vos WM. Segers A, et al. Microbiome Res Rep. 2023 Mar 13;2(1):6. doi: 10.20517/mrr.2023.05. eCollection 2023. Microbiome Res Rep. 2023. PMID: 38045608 Free PMC article. Review. - Peptidoglycan deacetylation controls type IV secretion and the intracellular survival of the bacterial pathogen Legionella pneumophila.
Boamah D, Gilmore MC, Bourget S, Ghosh A, Hossain MJ, Vogel JP, Cava F, O'Connor TJ. Boamah D, et al. Proc Natl Acad Sci U S A. 2023 Jun 6;120(23):e2119658120. doi: 10.1073/pnas.2119658120. Epub 2023 May 30. Proc Natl Acad Sci U S A. 2023. PMID: 37252954 Free PMC article. - Microbiological Quality of Raw Donkey Milk from Serbia and Its Antibacterial Properties at Pre-Cooling Temperature.
Šarić L, Premović T, Šarić B, Čabarkapa I, Todorić O, Miljanić J, Lazarević J, Karabasil N. Šarić L, et al. Animals (Basel). 2023 Jan 17;13(3):327. doi: 10.3390/ani13030327. Animals (Basel). 2023. PMID: 36766215 Free PMC article.
References
- Kocks C, Gouin E, Tabouret M, Berche P, Ohayon H, Cossart P. Cell. 1992;68:521–531. - PubMed
- Gaillard JL, Berche P, Frehel C, Gouin E, Cossart P. Cell. 1991;65:1127–1141. - PubMed
- Lebrun M, Mengaud J, Ohayon H, Nato F, Cossart P. Mol Microbiol. 1996;21:579–592. - PubMed
- Dramsi S, Biswas I, Maguin E, Braun L, Mastroeni P, Cossart P. Mol Microbiol. 1995;16:251–261. - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Medical
Molecular Biology Databases