Helicobacter pylori Peptidoglycan Modifications Confer Lysozyme Resistance and Contribute to Survival in the Host

Wang, Ge; Lo, Leja F.; Forsberg, Lennart S.; Maier, Robert J.

doi:10.1128/mbio.00409-12

Helicobacter pylori Peptidoglycan Modifications Confer Lysozyme Resistance and Contribute to Survival in the Host

Journal Article · Mon Dec 03 23:00:00 EST 2012 · mBio (Online)

DOI:https://doi.org/10.1128/mbio.00409-12· OSTI ID:1626104

Wang, Ge ^[1]; Lo, Leja F. ^[2]; Forsberg, Lennart S. ^[3]; Maier, Robert J. ^[2]

Univ. of Georgia, Athens, GA (United States). Dept. of Microbiology; DOE/OSTI
Univ. of Georgia, Athens, GA (United States). Dept. of Microbiology
Univ. of Georgia, Athens, GA (United States). The Complex Carbohydrate Research Center

The prominent host muramidase lysozyme cleaves bacterial peptidoglycan (PG), and the enzyme is abundant in mucosal secretions. The lytic enzyme susceptibility of Gram-negative bacteria and mechanisms they use to thwart lytic enzyme activity are poorly studied. We previously characterized a Helicobacter pylori PG modification enzyme, an N-deacetylase (PgdA) involved in lysozyme resistance. In this study, another PG modification enzyme, a putative PG O-acetyltransferase (PatA), was identified. Mass spectral analysis of the purified PG demonstrated that a patA strain contained a greatly reduced amount of acetylated muropeptides, indicating a role for PatA in H. pylori PG O-acetylation. The PG modification mutant strains (pgdA, patA, or pgdA patA) were more susceptible to lysozyme killing than the parent, but this assay required high lysozyme levels (up to 50 mg/ml). However, addition of host lactoferrin conferred lysozyme sensitivity to H. pylori, at physiologically relevant concentrations of both host components (3 mg/ml lactoferrin plus 0.3 mg/ml lysozyme). The pgdA patA double mutant strain was far more susceptible to lysozyme/lactoferrin killing than the parent. Peptidoglycan purified from a pgdA patA mutant was five times more sensitive to lysozyme than PG from the parent strain, while PG from both single mutants displayed intermediate sensitivity. Both sensitivity assays for whole cells and for purified PGs indicated that the modifications mediated by PgdA and PatA have a synergistic effect, conferring lysozyme tolerance. In a mouse infection model, significant colonization deficiency was observed for the double mutant at 3 weeks postinoculation. The results show that PG modifications affect the survival of a Gram-negative pathogen.

View Accepted Manuscript (DOE)

Research Organization:: Univ. of Georgia, Athens, GA (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

Grant/Contract Number:: FG02-93ER20097

OSTI ID:: 1626104

Journal Information:: mBio (Online), Journal Name: mBio (Online) Journal Issue: 6 Vol. 3; ISSN 2150-7511

Publisher:: American Society for Microbiology (ASM)Copyright Statement

Country of Publication:: United States

Language:: English

References (51)

Ultrastructural Localization of Lysozyme in Human Neutrophils by Immunogold Cramer, Elisabeth M.; Breton-Gorius, Janine Journal of Leukocyte Biology, Vol. 41, Issue 3 https://doi.org/10.1002/jlb.41.3.242	journal	March 1987
Recognition of bacterial peptidoglycan by the innate immune system Dziarski, R. Cellular and Molecular Life Sciences (CMLS), Vol. 60, Issue 9 https://doi.org/10.1007/s00018-003-3019-6	journal	September 2003
O-Acetylated peptidoglycan: Controlling the activity of bacterial autolysins and lytic enzymes of innate immune systems Moynihan, Patrick J.; Clarke, Anthony J. The International Journal of Biochemistry & Cell Biology, Vol. 43, Issue 12 https://doi.org/10.1016/j.biocel.2011.08.007	journal	December 2011
Lactoferrin—a multifunctional protein with antimicrobial properties Farnaud, Sebastien; Evans, Robert W. Molecular Immunology, Vol. 40, Issue 7 https://doi.org/10.1016/s0161-5890(03)00152-4	journal	November 2003
A critical role for peptidoglycan N-deacetylation in Listeria evasion from the host innate immune system Boneca, Ivo G.; Dussurget, Olivier; Cabanes, Didier Proceedings of the National Academy of Sciences, Vol. 104, Issue 3 https://doi.org/10.1073/pnas.0609672104	journal	January 2007
Oxidative Stress-induced Peptidoglycan Deacetylase in Helicobacter pylori Wang, Ge; Olczak, Adriana; Forsberg, Lennart S. Journal of Biological Chemistry, Vol. 284, Issue 11 https://doi.org/10.1074/jbc.m808071200	journal	March 2009
Lysozyme is an inducible marker of macrophage activation in murine tissues as demonstrated by in situ hybridization. Keshav, S.; Chung, P.; Milon, G. Journal of Experimental Medicine, Vol. 174, Issue 5 https://doi.org/10.1084/jem.174.5.1049	journal	November 1991
Nod1 Detects a Unique Muropeptide from Gram-Negative Bacterial Peptidoglycan Girardin, S. E. Science, Vol. 300, Issue 5625 https://doi.org/10.1126/science.1084677	journal	June 2003
Helicobacter pylori. Dunn, B. E.; Cohen, H.; Blaser, M. J. Clinical Microbiology Reviews, Vol. 10, Issue 4 https://doi.org/10.1128/cmr.10.4.720	journal	October 1997
Peptidoglycan Deacetylation in Helicobacter pylori Contributes to Bacterial Survival by Mitigating Host Immune Responses Wang, Ge; Maier, Susan E.; Lo, Leja F. Infection and Immunity, Vol. 78, Issue 11 https://doi.org/10.1128/iai.00307-10	journal	August 2010
Modifications to the Peptidoglycan Backbone Help Bacteria To Establish Infection Davis, Kimberly M.; Weiser, Jeffrey N. Infection and Immunity, Vol. 79, Issue 2 https://doi.org/10.1128/iai.00651-10	journal	November 2010
Characterization of Helicobacter pylori Lytic Transglycosylases Slt and MltD Chaput, C.; Labigne, A.; Boneca, I. G. Journal of Bacteriology, Vol. 189, Issue 2 https://doi.org/10.1128/jb.01270-06	journal	November 2006
The Morphological Transition of Helicobacter pyloriCells from Spiral to Coccoid Is Preceded by a Substantial Modification of the Cell Wall Costa, Katyssulla; Bacher, Gerold; Allmaier, Günter Journal of Bacteriology, Vol. 181, Issue 12 https://doi.org/10.1128/jb.181.12.3710-3715.1999	journal	June 1999
Role of AmiA in the Morphological Transition of Helicobacter pylori and in Immune Escape Chaput, Catherine; Ecobichon, Chantal; Cayet, Nadège PLoS Pathogens, Vol. 2, Issue 9 https://doi.org/10.1371/journal.ppat.0020097	journal	September 2006
A New Family of Lysozyme Inhibitors Contributing to Lysozyme Tolerance in Gram-Negative Bacteria Callewaert, Lien; Aertsen, Abram; Deckers, Daphne PLoS Pathogens, Vol. 4, Issue 3 https://doi.org/10.1371/journal.ppat.1000019	journal	March 2008
Resistance to Mucosal Lysozyme Compensates for the Fitness Deficit of Peptidoglycan Modifications by Streptococcus pneumoniae Davis, Kimberly M.; Akinbi, Henry T.; Standish, Alistair J. PLoS Pathogens, Vol. 4, Issue 12 https://doi.org/10.1371/journal.ppat.1000241	journal	December 2008
Multiple Peptidoglycan Modification Networks Modulate Helicobacter pylori's Cell Shape, Motility, and Colonization Potential Sycuro, Laura K.; Wyckoff, Timna J.; Biboy, Jacob PLoS Pathogens, Vol. 8, Issue 3 https://doi.org/10.1371/journal.ppat.1002603	journal	March 2012
Periplasmic lysozyme inhibitor contributes to lysozyme resistance in Escherichia coli Deckers, D.; Masschalck, B.; Aertsen, A. Cellular and Molecular Life Sciences (CMLS), Vol. 61, Issue 10 https://doi.org/10.1007/s00018-004-4066-3	journal	May 2004
Lysozymes in the animal kingdom Callewaert, Lien; Michiels, Chris W. Journal of Biosciences, Vol. 35, Issue 1 https://doi.org/10.1007/s12038-010-0015-5	journal	February 2010
Lactoferrin—a multifunctional protein with antimicrobial properties Farnaud, Sebastien; Evans, Robert W. Molecular Immunology, Vol. 40, Issue 7 https://doi.org/10.1016/S0161-5890(03)00152-4	journal	November 2003
Peptidoglycan Crosslinking Relaxation Promotes Helicobacter pylori's Helical Shape and Stomach Colonization Sycuro, Laura K.; Pincus, Zachary; Gutierrez, Kimberley D. Cell, Vol. 141, Issue 5 https://doi.org/10.1016/j.cell.2010.03.046	journal	May 2010
The complete genome sequence of the gastric pathogen Helicobacter pylori Tomb, Jean-F.; White, Owen; Kerlavage, Anthony R. Nature, Vol. 388, Issue 6642 https://doi.org/10.1038/41483	journal	August 1997
Nod1 responds to peptidoglycan delivered by the Helicobacter pylori cag pathogenicity island Viala, Jérôme; Chaput, Catherine; Boneca, Ivo G. Nature Immunology, Vol. 5, Issue 11 https://doi.org/10.1038/ni1131	journal	October 2004
Increased expression of antimicrobial peptides and lysozyme in colonic epithelial cells of patients with ulcerative colitis: Antimicrobial peptides in inflamed intestinal epithelium Fahlgren, A.; HammarstrÖM, S.; Danielsson, Å. Clinical & Experimental Immunology, Vol. 131, Issue 1 https://doi.org/10.1046/j.1365-2249.2003.02035.x	journal	January 2003
O -Acetylation of Peptidoglycan in Gram-negative Bacteria : IDENTIFICATION AND CHARACTERIZATION OF PEPTIDOGLYCAN O-ACETYLTRANSFERASE IN NEISSERIA GONORRHOEAE Moynihan, Patrick J.; Clarke, Anthony J. Journal of Biological Chemistry, Vol. 285, Issue 17 https://doi.org/10.1074/jbc.M110.107086	journal	February 2010
Peptidoglycan N-Acetylglucosamine Deacetylases from Bacillus cereus, Highly Conserved Proteins in Bacillus anthracis Psylinakis, Emmanuel; Boneca, Ivo G.; Mavromatis, Konstantinos Journal of Biological Chemistry, Vol. 280, Issue 35 https://doi.org/10.1074/jbc.M407426200	journal	September 2005
Oxidative Stress-induced Peptidoglycan Deacetylase in Helicobacter pylori Wang, Ge; Olczak, Adriana; Forsberg, Lennart S. Journal of Biological Chemistry, Vol. 284, Issue 11 https://doi.org/10.1074/jbc.M808071200	journal	March 2009
The pgdA Gene Encodes for a PeptidoglycanN-Acetylglucosamine Deacetylase in Streptococcus pneumoniae Vollmer, Waldemar; Tomasz, Alexander Journal of Biological Chemistry, Vol. 275, Issue 27 https://doi.org/10.1074/jbc.M910189199	journal	July 2000
Helicobacter pylori Moss, Steven F.; Sood, Shivani Current Opinion in Infectious Diseases, Vol. 16, Issue 5 https://doi.org/10.1097/00001432-200310000-00011	journal	January 2003
Peptidoglycan N-acetylglucosamine deacetylation decreases autolysis in Lactococcus lactis Meyrand, Mickael; Boughammoura, Aïda; Courtin, Pascal Microbiology, Vol. 153, Issue 10 https://doi.org/10.1099/mic.0.2007/005835-0	journal	October 2007
Helicobacter pylori tissue tropism: mouse-colonizing strains can target different gastric niches Akada, Junko K.; Ogura, Keiji; Dailidiene, Daiva Microbiology, Vol. 149, Issue 7 https://doi.org/10.1099/mic.0.26129-0	journal	July 2003
Pathogenesis of Helicobacter pylori Infection Sgouras, Dionyssios N.; Trang, Tran Thi Huyen; Yamaoka, Yoshio Helicobacter, Vol. 20 https://doi.org/10.1111/hel.12251	journal	September 2015
Pathogenesis of Helicobacter pylori infection Camilo, Vania; Sugiyama, Toshiro; Touati, Eliette Helicobacter, Vol. 22 https://doi.org/10.1111/hel.12405	journal	September 2017
Why are pathogenic staphylococci so lysozyme resistant? The peptidoglycan O-acetyltransferase OatA is the major determinant for lysozyme resistance of Staphylococcus aureus: Lysozyme resistance of S. aureus Bera, Agnieszka; Herbert, Silvia; Jakob, Andreas Molecular Microbiology, Vol. 55, Issue 3 https://doi.org/10.1111/j.1365-2958.2004.04446.x	journal	December 2004
A M23B family metallopeptidase of Helicobacter pylori required for cell shape, pole formation and virulence: M23 peptidase required for cell shape Bonis, Mathilde; Ecobichon, Chantal; Guadagnini, Stephanie Molecular Microbiology, Vol. 78, Issue 4 https://doi.org/10.1111/j.1365-2958.2010.07383.x	journal	September 2010
Bacterial membrane vesicles deliver peptidoglycan to NOD1 in epithelial cells Kaparakis, Maria; Turnbull, Lynne; Carneiro, Leticia Cellular Microbiology, Vol. 12, Issue 3 https://doi.org/10.1111/j.1462-5822.2009.01404.x	journal	March 2010
Pathogenesis of Helicobacter pylori Infection: H. pylori Pathogenesis Basso, Daniela; Plebani, Mario; Kusters, Johannes G. Helicobacter, Vol. 15 https://doi.org/10.1111/j.1523-5378.2010.00781.x	journal	September 2010
Structural variation in the glycan strands of bacterial peptidoglycan Vollmer, Waldemar FEMS Microbiology Reviews, Vol. 32, Issue 2 https://doi.org/10.1111/j.1574-6976.2007.00088.x	journal	March 2008
Pathogenesis of Helicobacter pylori Infection Kusters, J. G.; van Vliet, A. H. M.; Kuipers, E. J. Clinical Microbiology Reviews, Vol. 19, Issue 3 https://doi.org/10.1128/CMR.00054-05	journal	July 2006
Helicobacter pylori. Dunn, B. E.; Cohen, H.; Blaser, M. J. Clinical Microbiology Reviews, Vol. 10, Issue 4 https://doi.org/10.1128/CMR.10.4.720	journal	October 1997
Peptidoglycan Deacetylation in Helicobacter pylori Contributes to Bacterial Survival by Mitigating Host Immune Responses Wang, Ge; Maier, Susan E.; Lo, Leja F. Infection and Immunity, Vol. 78, Issue 11 https://doi.org/10.1128/IAI.00307-10	journal	August 2010
Enterococcus faecalis Constitutes an Unusual Bacterial Model in Lysozyme Resistance Hebert, L.; Courtin, P.; Torelli, R. Infection and Immunity, Vol. 75, Issue 11 https://doi.org/10.1128/IAI.00571-07	journal	September 2007
Modifications to the Peptidoglycan Backbone Help Bacteria To Establish Infection Davis, Kimberly M.; Weiser, Jeffrey N. Infection and Immunity, Vol. 79, Issue 2 https://doi.org/10.1128/IAI.00651-10	journal	November 2010
A RecB-Like Helicase in Helicobacter pylori Is Important for DNA Repair and Host Colonization Wang, Ge; Maier, Robert J. Infection and Immunity, Vol. 77, Issue 1 https://doi.org/10.1128/IAI.00970-08	journal	November 2008
Dual Roles of Helicobacter pylori NapA in Inducing and Combating Oxidative Stress Wang, Ge; Hong, Yang; Olczak, Adriana Infection and Immunity, Vol. 74, Issue 12 https://doi.org/10.1128/IAI.00991-06	journal	October 2006
Mutations Affecting Peptidoglycan Acetylation in Neisseria gonorrhoeae and Neisseria meningitidis Dillard, Joseph P.; Hackett, Kathleen T. Infection and Immunity, Vol. 73, Issue 9 https://doi.org/10.1128/IAI.73.9.5697-5705.2005	journal	September 2005
Characterization of Helicobacter pylori Lytic Transglycosylases Slt and MltD Chaput, C.; Labigne, A.; Boneca, I. G. Journal of Bacteriology, Vol. 189, Issue 2 https://doi.org/10.1128/JB.01270-06	journal	November 2006
The Morphological Transition of Helicobacter pyloriCells from Spiral to Coccoid Is Preceded by a Substantial Modification of the Cell Wall Costa, Katyssulla; Bacher, Gerold; Allmaier, Günter Journal of Bacteriology, Vol. 181, Issue 12 https://doi.org/10.1128/JB.181.12.3710-3715.1999	journal	June 1999
Helicobacter pylori Moss, S. Gut, Vol. 41, Issue 5 https://doi.org/10.1136/gut.41.5.e723	journal	November 1997
Killing of gram-negative bacteria by lactoferrin and lysozyme. Ellison, R. T.; Giehl, T. J. Journal of Clinical Investigation, Vol. 88, Issue 4 https://doi.org/10.1172/JCI115407	journal	October 1991
NOD1-Mediated Mucosal Host Defense against Helicobacter pylori Watanabe, Tomohiro; Asano, Naoki; Kitani, Atsushi International Journal of Inflammation, Vol. 2010 https://doi.org/10.4061/2010/476482	journal	January 2010

Cited By (19)

Comparative genome analysis of Weissella ceti, an emerging pathogen of farm-raised rainbow trout Figueiredo, Henrique C. P.; Soares, Siomar C.; Pereira, Felipe L. BMC Genomics, Vol. 16, Issue 1 https://doi.org/10.1186/s12864-015-2324-4	journal	December 2015
Biochemical Characterization of Hypothetical Proteins from Helicobacter pylori Choi, Han-Pil; Juarez, Silvia; Ciordia, Sergio PLoS ONE, Vol. 8, Issue 6 https://doi.org/10.1371/journal.pone.0066605	journal	June 2013
Helicobacter pylori cag Pathogenicity Island's Role in B7-H1 Induction and Immune Evasion Lina, Taslima T.; Alzahrani, Shatha; House, Jennifer PLOS ONE, Vol. 10, Issue 3 https://doi.org/10.1371/journal.pone.0121841	journal	March 2015
Biochemical characterization of the Helicobacter pylori bactofilin-homolog HP1542 Holtrup, Sven; Heimerl, Thomas; Linne, Uwe PLOS ONE, Vol. 14, Issue 6 https://doi.org/10.1371/journal.pone.0218474	journal	June 2019
In vitro characterization of the antivirulence target of Gram-positive pathogens, peptidoglycan O-acetyltransferase A (OatA) Sychantha, David; Jones, Carys S.; Little, Dustin J. PLOS Pathogens, Vol. 13, Issue 10 https://doi.org/10.1371/journal.ppat.1006667	journal	October 2017
Mechanistic Pathways for Peptidoglycan O-Acetylation and De-O-Acetylation Sychantha, David; Brott, Ashley S.; Jones, Carys S. Frontiers in Microbiology, Vol. 9 https://doi.org/10.3389/fmicb.2018.02332	journal	October 2018
Comparative Analysis of Peptidoglycans From Pseudomonas aeruginosa Isolates Recovered From Chronic and Acute Infections Torrens, Gabriel; Escobar-Salom, María; Pol-Pol, Elisabet Frontiers in Microbiology, Vol. 10 https://doi.org/10.3389/fmicb.2019.01868	journal	August 2019
Human Lysozyme Synergistically Enhances Bactericidal Dynamics and Lowers the Resistant Mutant Prevention Concentration for Metronidazole to Helicobacter pylori by Increasing Cell Permeability Zhang, Xiaolin; Jiang, Anmin; Yu, Hao Molecules, Vol. 21, Issue 11 https://doi.org/10.3390/molecules21111435	journal	October 2016
Morphology of Helicobacter pylori as a result of peptidoglycan and cytoskeleton rearrangements Krzyżek, Paweł; Gościniak, Grażyna Gastroenterology Review, Vol. 13, Issue 3 https://doi.org/10.5114/pg.2018.78284	journal	January 2018
Analysis of carbohydrates and glycoconjugates by matrix-assisted laser desorption/ionization mass spectrometry: An update for 2011-2012: ANALYSIS OF CARBOHYDRATES AND GLYCOCONJUGATES Harvey, David J. Mass Spectrometry Reviews, Vol. 36, Issue 3 https://doi.org/10.1002/mas.21471	journal	August 2015
Identification of novel Legionella genes required for endosymbiosis in Paramecium based on comparative genome analysis with Holospora spp. Watanabe, Kenta; Suzuki, Haruo; Nishida, Takashi FEMS Microbiology Ecology, Vol. 94, Issue 11 https://doi.org/10.1093/femsec/fiy162	journal	August 2018
Peptidoglycan recycling contributes to intrinsic resistance to fosfomycin in Acinetobacter baumannii Gil-Marqués, María Luisa; Moreno-Martínez, Patricia; Costas, Coloma Journal of Antimicrobial Chemotherapy, Vol. 73, Issue 11 https://doi.org/10.1093/jac/dky289	journal	August 2018
A novel peptidoglycan D,L-endopeptidase induced by S almonella inside eukaryotic cells contributes to virulence: Peptidoglycan of intracellular Salmonella Rico-Pérez, Gadea; Pezza, Alejandro; Pucciarelli, M. Graciela Molecular Microbiology, Vol. 99, Issue 3 https://doi.org/10.1111/mmi.13248	journal	November 2015
Aconitase-Mediated Posttranscriptional Regulation of Helicobacter pylori Peptidoglycan Deacetylase Austin, C. M.; Maier, R. J. Journal of Bacteriology, Vol. 195, Issue 23 https://doi.org/10.1128/jb.00720-13	journal	September 2013
Hydrogen Metabolism in Helicobacter pylori Plays a Role in Gastric Carcinogenesis through Facilitating CagA Translocation Wang, Ge; Romero-Gallo, Judith; Benoit, Stéphane L. mBio, Vol. 7, Issue 4 https://doi.org/10.1128/mbio.01022-16	journal	August 2016
Interplay between Peptidoglycan Biology and Virulence in Gram-Negative Pathogens Juan, Carlos; Torrens, Gabriel; Barceló, Isabel Maria Microbiology and Molecular Biology Reviews, Vol. 82, Issue 4 https://doi.org/10.1128/mmbr.00033-18	journal	September 2018
From bacterial killing to immune modulation: Recent insights into the functions of lysozyme Ragland, Stephanie A.; Criss, Alison K. PLOS Pathogens, Vol. 13, Issue 9 https://doi.org/10.1371/journal.ppat.1006512	journal	September 2017
Development of a High Throughput Screen for the Identification of Inhibitors of Peptidoglycan O-Acetyltransferases, New Potential Antibacterial Targets Brott, Ashley S.; Jones, Carys S.; Clarke, Anthony J. Antibiotics, Vol. 8, Issue 2 https://doi.org/10.3390/antibiotics8020065	journal	May 2019
Peptidoglycan O-Acetylation as a Virulence Factor: Its Effect on Lysozyme in the Innate Immune System Brott, Ashley S.; Clarke, Anthony J. Antibiotics, Vol. 8, Issue 3 https://doi.org/10.3390/antibiotics8030094	journal	July 2019

Similar Records

Modification of Helicobacter pylori peptidoglycan enhances NOD1 activation and promotes cancer of the stomach

Journal Article · Sun Mar 01 23:00:00 EST 2015 · Cancer Research · OSTI ID:1345502

Cloning, expression, and characterization of a peptidoglycan hydrolase from the Burkholderia pseudomallei phage ST79

Journal Article · Thu Sep 15 00:00:00 EDT 2016 · AMB Express · OSTI ID:1627035

In vitro characterization of the antivirulence target of Gram-positive pathogens, peptidoglycan O-acetyltransferase A (OatA)

Journal Article · Fri Oct 27 00:00:00 EDT 2017 · PLoS Pathogens · OSTI ID:1425067

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
Microbiology

Helicobacter pylori Peptidoglycan Modifications Confer Lysozyme Resistance and Contribute to Survival in the Host

Citation Formats

References (51)

Cited By (19)

Similar Records

Related Subjects