{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,10]],"date-time":"2026-04-10T07:57:40Z","timestamp":1775807860088,"version":"3.50.1"},"reference-count":42,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2020,4,16]],"date-time":"2020-04-16T00:00:00Z","timestamp":1586995200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100008530","name":"European Regional Development Fund","doi-asserted-by":"publisher","award":["NORTE-01-0145-FEDER-030020 PTDC\/SAU-INF\/30020\/2017"],"award-info":[{"award-number":["NORTE-01-0145-FEDER-030020 PTDC\/SAU-INF\/30020\/2017"]}],"id":[{"id":"10.13039\/501100008530","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["CEEC-Institutional 2017 program"],"award-info":[{"award-number":["CEEC-Institutional 2017 program"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>The cell wall of Listeria monocytogenes (Lm), a major intracellular foodborne bacterial pathogen, comprises a thick peptidoglycan layer that serves as a scaffold for glycopolymers such as wall teichoic acids (WTAs). WTAs contain non-essential sugar substituents whose absence prevents bacteriophage binding and impacts antigenicity, sensitivity to antimicrobials, and virulence. Here, we demonstrated, for the first time, the triple function of Lm WTA glycosylations in the following: (1) supporting the correct anchoring of major Lm virulence factors at the bacterial surface, namely Ami and InlB; (2) promoting Lm resistance to antimicrobial peptides (AMPs); and (3) decreasing Lm sensitivity to some antibiotics. We showed that while the decoration of WTAs by rhamnose in Lm serovar 1\/2a and by galactose in serovar 4b are important for the surface anchoring of Ami and InlB, N-acetylglucosamine in serovar 1\/2a and glucose in serovar 4b are dispensable for the surface association of InlB or InlB\/Ami. We found that the absence of a single glycosylation only had a slight impact on the sensibility of Lm to AMPs and antibiotics, however the concomitant deficiency of both glycosylations (rhamnose and N-acetylglucosamine in serovar 1\/2a, and galactose and glucose in serovar 4b) significantly impaired the Lm capacity to overcome the action of antimicrobials. We propose WTA glycosylation as a broad mechanism used by Lm, not only to properly anchor surface virulence factors, but also to resist AMPs and antibiotics. WTA glycosyltransferases thus emerge as promising drug targets to attenuate the virulence of bacterial pathogens, while increasing their susceptibility to host immune defenses and potentiating the action of antibiotics.<\/jats:p>","DOI":"10.3390\/pathogens9040290","type":"journal-article","created":{"date-parts":[[2020,4,16]],"date-time":"2020-04-16T13:01:39Z","timestamp":1587042099000},"page":"290","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":33,"title":["Listeria monocytogenes Wall Teichoic Acid Glycosylation Promotes Surface Anchoring of Virulence Factors, Resistance to Antimicrobial Peptides, and Decreased Susceptibility to Antibiotics"],"prefix":"10.3390","volume":"9","author":[{"given":"Diana","family":"Meireles","sequence":"first","affiliation":[{"name":"i3S\u2013Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Group of Molecular Microbiology, IBMC\u2013Instituto de Biologia Celular e Molecular, 4200-135 Porto, Portugal"}]},{"given":"Rita","family":"Pombinho","sequence":"additional","affiliation":[{"name":"i3S\u2013Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Group of Molecular Microbiology, IBMC\u2013Instituto de Biologia Celular e Molecular, 4200-135 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6828-1772","authenticated-orcid":false,"given":"Filipe","family":"Carvalho","sequence":"additional","affiliation":[{"name":"i3S\u2013Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Group of Molecular Microbiology, IBMC\u2013Instituto de Biologia Celular e Molecular, 4200-135 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8578-0461","authenticated-orcid":false,"given":"Sandra","family":"Sousa","sequence":"additional","affiliation":[{"name":"i3S\u2013Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Cell Biology of Bacterial Infections Group, IBMC\u2013Instituto de Biologia Molecular e Celular, 4200-135 Porto, Portugal"}]},{"given":"Didier","family":"Cabanes","sequence":"additional","affiliation":[{"name":"i3S\u2013Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Group of Molecular Microbiology, IBMC\u2013Instituto de Biologia Celular e Molecular, 4200-135 Porto, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2020,4,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"19484","DOI":"10.1073\/pnas.1112371108","article-title":"Illuminating the landscape of host-pathogen interactions with the bacterium Listeria monocytogenes","volume":"108","author":"Cossart","year":"2011","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"European Food Safety Authority, and European Centre for Disease Prevention and Control (2017). The European Union summary report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2016. EFSA J., 15, e05077.","DOI":"10.2903\/j.efsa.2017.5077"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"48","DOI":"10.3389\/fcimb.2014.00048","article-title":"How Listeria monocytogenes organizes its surface for virulence","volume":"4","author":"Carvalho","year":"2014","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1038\/nrmicro1861","article-title":"Teichoic acids and related cell-wall glycopolymers in Gram-positive physiology and host interactions","volume":"6","author":"Weidenmaier","year":"2008","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1146\/annurev-micro-091213-112949","article-title":"Lipoteichoic acid synthesis and function in gram-positive bacteria","volume":"68","author":"Percy","year":"2014","journal-title":"Annu. Rev. Microbiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1146\/annurev-micro-092412-155620","article-title":"Wall teichoic acids of gram-positive bacteria","volume":"67","author":"Brown","year":"2013","journal-title":"Annu. Rev. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"S92","DOI":"10.1007\/BF01639729","article-title":"Biochemistry of the cell surface of Listeria strains: A locating general view","volume":"16","author":"Fiedler","year":"1988","journal-title":"Infection"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1128\/jb.168.1.115-122.1986","article-title":"Structural studies on lipoteichoic acids from four Listeria strains","volume":"168","author":"Uchikawa","year":"1986","journal-title":"J. Bacteriol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2054","DOI":"10.1128\/IAI.69.4.2054-2065.2001","article-title":"Identification of new genes involved in the virulence of Listeria monocytogenes by signature-tagged transposon mutagenesis","volume":"69","author":"Autret","year":"2001","journal-title":"Infect. Immun."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1419","DOI":"10.1111\/j.1365-2958.2011.07774.x","article-title":"The cell wall binding domain of Listeria bacteriophage endolysin PlyP35 recognizes terminal GlcNAc residues in cell wall teichoic acid","volume":"81","author":"Eugster","year":"2011","journal-title":"Mol. Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1089\/fpd.2008.0154","article-title":"The role of L. monocytogenes serotype 4b gtcA in gastrointestinal listeriosis in A\/J mice","volume":"6","author":"Faith","year":"2009","journal-title":"Foodborne Pathog. Dis."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"418","DOI":"10.1128\/JB.181.2.418-425.1999","article-title":"Cell wall teichoic acid glycosylation in Listeria monocytogenes serotype 4b requires gtcA, a novel, serogroup-specific gene","volume":"181","author":"Promadej","year":"1999","journal-title":"J. Bacteriol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"714","DOI":"10.1111\/mmi.13353","article-title":"Listeria monocytogenes wall teichoic acid decoration in virulence and cell-to-cell spread","volume":"101","author":"Spears","year":"2016","journal-title":"Mol. Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1653","DOI":"10.1128\/AEM.01773-07","article-title":"Teichoic acid glycosylation mediated by gtcA is required for phage adsorption and susceptibility of Listeria monocytogenes serotype 4b","volume":"74","author":"Cheng","year":"2008","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1111\/mmi.13009","article-title":"Bacteriophage predation promotes serovar diversification in Listeria monocytogenes","volume":"97","author":"Eugster","year":"2015","journal-title":"Mol. Microbiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/j.ijmm.2010.05.002","article-title":"Listeria monocytogenes lineages: Genomics, evolution, ecology, and phenotypic characteristics","volume":"301","author":"Orsi","year":"2011","journal-title":"Int. J. Med. Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Carvalho, F., Atilano, M.L., Pombinho, R., Covas, G., Gallo, R.L., Filipe, S.R., Sousa, S., and Cabanes, D. (2015). L-Rhamnosylation of Listeria monocytogenes Wall Teichoic Acids Promotes Resistance to Antimicrobial Peptides by Delaying Interaction with the Membrane. PLoS Pathog., 11.","DOI":"10.1371\/journal.ppat.1004919"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1046\/j.1365-2958.1997.4621825.x","article-title":"InlB: An invasion protein of Listeria monocytogenes with a novel type of surface association","volume":"25","author":"Braun","year":"1997","journal-title":"Mol. Microbiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1212","DOI":"10.1111\/j.1365-2958.2001.02208.x","article-title":"The Autolysin Ami contributes to the adhesion of Listeria monocytogenes to eucaryotic cells, via its cell wall anchor","volume":"39","author":"Milohanic","year":"2001","journal-title":"Mol. Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"902","DOI":"10.1046\/j.1365-2958.1999.01652.x","article-title":"Interaction between the protein InlB of Listeria monocytogenes and lipoteichoic acid: A novel mechanism of protein association at the surface of gram-positive bacteria","volume":"34","author":"Jonquieres","year":"1999","journal-title":"Mol. Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2029","DOI":"10.1128\/JB.00116-16","article-title":"Identification of a Lipoteichoic Acid Glycosyltransferase Enzyme Reveals that GW-Domain-Containing Proteins Can Be Retained in the Cell Wall of Listeria monocytogenes in the Absence of Lipoteichoic Acid or Its Modifications","volume":"198","author":"Percy","year":"2016","journal-title":"J. Bacteriol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3941","DOI":"10.1111\/1462-2920.14351","article-title":"l-Rhamnosylation of wall teichoic acids promotes efficient surface association of Listeria monocytogenes virulence factors InlB and Ami through interaction with GW domains","volume":"20","author":"Carvalho","year":"2018","journal-title":"Environ. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Sumrall, E.T., Shen, Y., Keller, A.P., Rismondo, J., Pavlou, M., Eugster, M.R., Boulos, S., Disson, O., Thouvenot, P., and Kilcher, S. (2019). Phage resistance at the cost of virulence: Listeria monocytogenes serovar 4b requires galactosylated teichoic acids for InlB-mediated invasion. PLoS Pathog., 15.","DOI":"10.1371\/journal.ppat.1008032"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"18909","DOI":"10.1073\/pnas.1209126109","article-title":"Methicillin resistance in Staphylococcus aureus requires glycosylated wall teichoic acids","volume":"109","author":"Brown","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_25","unstructured":"Schaechter, M., and Schaechter, M. (2009). Listeria monocytogenes. Encyclopedia of Microbiolog, Institut Pasteur."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"4283","DOI":"10.1038\/s41467-019-12072-1","article-title":"A hybrid sub-lineage of Listeria monocytogenes comprising hypervirulent isolates","volume":"10","author":"Yin","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"3942","DOI":"10.1111\/j.1742-4658.2011.08302.x","article-title":"Antimicrobial peptides important in innate immunity","volume":"278","author":"Cederlund","year":"2011","journal-title":"FEBS J."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Malanovic, N., and Lohner, K. (2016). Antimicrobial Peptides Targeting Gram-Positive Bacteria. Pharmaceuticals, 9.","DOI":"10.3390\/ph9030059"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Peters, B.M., Shirtliff, M.E., and Jabra-Rizk, M.A. (2010). Antimicrobial peptides: Primeval molecules or future drugs?. PLoS Pathog., 6.","DOI":"10.1371\/journal.ppat.1001067"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"238","DOI":"10.1038\/nrmicro1098","article-title":"Antimicrobial peptides: Pore formers or metabolic inhibitors in bacteria?","volume":"3","author":"Brogden","year":"2005","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1038\/nrmicro1441","article-title":"The co-evolution of host cationic antimicrobial peptides and microbial resistance","volume":"4","author":"Peschel","year":"2006","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1021\/cb300413m","article-title":"Inhibition of WTA synthesis blocks the cooperative action of PBPs and sensitizes MRSA to beta-lactams","volume":"8","author":"Farha","year":"2013","journal-title":"ACS Chem. Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"3797","DOI":"10.1128\/AAC.00187-12","article-title":"Dual roles of FmtA in Staphylococcus aureus cell wall biosynthesis and autolysis","volume":"56","author":"Qamar","year":"2012","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1016\/j.virol.2014.12.035","article-title":"Receptor binding proteins of Listeria monocytogenes bacteriophages A118 and P35 recognize serovar-specific teichoic acids","volume":"477","author":"Bielmann","year":"2015","journal-title":"Virology"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Dunne, M., Hupfeld, M., Klumpp, J., and Loessner, M.J. (2018). Molecular Basis of Bacterial Host Interactions by Gram-Positive Targeting Bacteriophages. Viruses, 10.","DOI":"10.3390\/v10080397"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1277","DOI":"10.1111\/1541-4337.12387","article-title":"Emergence of Antibiotic Resistance in Listeria monocytogenes Isolated from Food Products: A Comprehensive Review","volume":"17","author":"Olaimat","year":"2018","journal-title":"Compr Rev Food Sci Food Saf."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"347","DOI":"10.3201\/eid0804.010312","article-title":"Antimicrobial use and antimicrobial resistance: A population perspective","volume":"8","author":"Lipsitch","year":"2002","journal-title":"Emerg. Infect. Dis."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"300","DOI":"10.1038\/nrmicro3232","article-title":"Targeting virulence: Can we make evolution-proof drugs?","volume":"12","author":"Allen","year":"2014","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"6887","DOI":"10.1128\/AEM.70.11.6887-6891.2004","article-title":"New vector for efficient allelic replacement in naturally nontransformable, low-GC-content, gram-positive bacteria","volume":"70","author":"Arnaud","year":"2004","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1468","DOI":"10.1093\/infdis\/jix118","article-title":"Listeria monocytogenes CadC regulates cadmium efflux and fine-tunes lipoprotein localization to escape the host immune response and promote infection","volume":"215","author":"Pombinho","year":"2017","journal-title":"J. Infect. Dis."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"849","DOI":"10.1126\/science.1063447","article-title":"Comparative genomics of Listeria species","volume":"294","author":"Glaser","year":"2001","journal-title":"Science"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1046\/j.1365-2958.1999.01560.x","article-title":"The 213-amino-acid leucine-rich repeat region of the Listeria monocytogenes InlB protein is sufficient for entry into mammalian cells, stimulation of PI 3-kinase and membrane ruffling","volume":"34","author":"Braun","year":"1999","journal-title":"Mol. Microbiol."}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/9\/4\/290\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T13:21:18Z","timestamp":1760361678000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/9\/4\/290"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,4,16]]},"references-count":42,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2020,4]]}},"alternative-id":["pathogens9040290"],"URL":"https:\/\/doi.org\/10.3390\/pathogens9040290","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,4,16]]}}}