{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T22:43:51Z","timestamp":1790981031219,"version":"4.1.0"},"reference-count":54,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,10,30]],"date-time":"2023-10-30T00:00:00Z","timestamp":1698624000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>The <jats:italic>dlt<\/jats:italic> operon encodes proteins responsible for the esterification of positively charged D-alanine on the wall teichoic acids and lipoteichoic acids of Gram-positive bacteria. This structural modification of the bacterial anionic surface in several species has been described to alter the physicochemical properties of the cell-wall. In addition, it has been linked to reduced sensibilities to cationic antimicrobial peptides and antibiotics.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We studied the D-alanylation of <jats:italic>Clostridioides difficile<\/jats:italic> polysaccharides with a complete deletion of the <jats:italic>dltDABC<\/jats:italic>operon in the 630 strain. To look for D-alanylation location, surface polysaccharides were purified and analyzed by NMR. Properties of the <jats:italic>dltDABC<\/jats:italic>mutant and the parental strains, were determined for bacterial surface\u2019s hydrophobicity, motility, adhesion, antibiotic resistance.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>We first confirmed the role of the <jats:italic>dltDABC<\/jats:italic>operon in D-alanylation. Then, we established the exclusive esterification of D-alanine on <jats:italic>C. difficile<\/jats:italic> lipoteichoic acid. Our data also suggest that D-alanylation modifies the cell-wall\u2019s properties, affecting the bacterial surface\u2019s hydrophobicity, motility, adhesion to biotic and abiotic surfaces,and biofilm formation. In addition, our mutant exhibitedincreased sensibilities to antibiotics linked to the membrane, especially bacitracin. A specific inhibitor DLT-1 of DltA reduces the D-alanylation rate in <jats:italic>C. difficile<\/jats:italic> but the inhibition was not sufficient to decrease the antibiotic resistance against bacitracin and vancomycin.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Our results suggest the D-alanylation of <jats:italic>C. difficile<\/jats:italic> as an interesting target to tackle <jats:italic>C. difficile<\/jats:italic> infections.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fmicb.2023.1267662","type":"journal-article","created":{"date-parts":[[2023,10,30]],"date-time":"2023-10-30T09:46:20Z","timestamp":1698659180000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":7,"title":["The absence of surface D-alanylation, localized on lipoteichoic acid, impacts the Clostridioides difficile way of life and antibiotic resistance"],"prefix":"10.3389","volume":"14","author":[{"given":"Pierre-Alexandre","family":"Lacotte","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sandrine","family":"Denis-Quanquin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva","family":"Chatonnat","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julie","family":"Le Bris","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Leparfait","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thierry","family":"Lequeux","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabelle","family":"Martin-Verstraete","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Candela","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,10,30]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1046\/j.1365-2958.2002.02723.x","article-title":"Formation of D-alanyl-lipoteichoic acid is required for adhesion and virulence of Listeria monocytogenes","volume":"43","author":"Abachin","year":"2002","journal-title":"Mol Microbiol."},{"key":"ref2","doi-asserted-by":"publisher","first-page":"86","DOI":"10.1016\/j.mib.2022.01.007","article-title":"Surface-displayed glycopolymers of Clostridioides difficile","volume":"66","author":"Anwar","year":"2022","journal-title":"Current Opin. 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