{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T05:08:39Z","timestamp":1789103319101,"version":"build-2803163510"},"reference-count":36,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T00:00:00Z","timestamp":1788998400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T00:00:00Z","timestamp":1788998400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100003246","name":"Nederlandse Organisatie voor Wetenschappelijk Onderzoek","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100003246","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001833","name":"Vrije Universiteit Amsterdam","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001833","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Molecular Microbiology"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Lactobacillus crispatus<\/jats:italic>\n                    <\/jats:styled-content>\n                    dominance in the vaginal microbiome is associated with beneficial health outcomes, yet strain\u2010level variation and its implications remain poorly understood. Here, we resolve the genomic context of three glycosyltransferase gene fragments (GT1\u20103) previously linked with dysbiotic states. Long\u2010read resequencing revealed that GT1\u20103 are part of a ~18.7\u2009kb Wzx\/Wzy\u2010dependent cell wall polysaccharide (CWPS) locus, containing several IS256\u2010family transposases. Serial propagation in\u00a0vitro produced isolates with 4.1\u2009kb excised via a composite transposon encompassing the GT3, UDP\u2010galactopyranose mutase, flippase, and hypothetical protein, demonstrating structural plasticity. Transmission electron microscopy showed a ~20%\u201325% thinner peptidoglycan layer in the derived strains, while FT\u2010IR and monosaccharide analysis indicated no gross changes. Molecular dynamics simulations suggest that GT3 contributes to the structural stability of the glycosyltransferase complex, without compromising catalytic function. Together, these findings establish the CWPS locus as a mobile, structurally plastic element that directly influences cell wall architecture in\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>L. crispatus<\/jats:italic>\n                    <\/jats:styled-content>\n                    .\n                  <\/jats:p>","DOI":"10.1111\/mmi.70112","type":"journal-article","created":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T04:47:38Z","timestamp":1789102058000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A Mobile Glycosylation Locus Modulates Cell Wall Architecture in\n                    <scp>\n                      <i>Lactobacillus crispatus<\/i>\n                    <\/scp>"],"prefix":"10.1111","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-6350-2129","authenticated-orcid":false,"given":"P.","family":"Achterberg","sequence":"first","affiliation":[{"name":"Amsterdam Institute for Life and Environment (A\u2010LIFE) Vrije Universiteit Amsterdam  Amsterdam the Netherlands"},{"name":"Department of Biotechnology Delft University of Technology  Delft the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-0262-5313","authenticated-orcid":false,"given":"M.","family":"Lekkerkerker","sequence":"additional","affiliation":[{"name":"Amsterdam Institute for Life and Environment (A\u2010LIFE) Vrije Universiteit Amsterdam  Amsterdam the Netherlands"},{"name":"Van't Hoff Institute for Molecular Sciences University of Amsterdam  Amsterdam the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"L.","family":"Koelemeijer","sequence":"additional","affiliation":[{"name":"Amsterdam Institute for Life and Environment (A\u2010LIFE) Vrije Universiteit Amsterdam  Amsterdam the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S.","family":"Illidge","sequence":"additional","affiliation":[{"name":"Amsterdam Institute for Life and Environment (A\u2010LIFE) Vrije Universiteit Amsterdam  Amsterdam the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J.","family":"Vroom","sequence":"additional","affiliation":[{"name":"Wageningen Electron Microscopy Centre Wageningen University &amp; Research  Wageningen the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6119-9450","authenticated-orcid":false,"given":"R. Y.","family":"Hertzberger","sequence":"additional","affiliation":[{"name":"Amsterdam Institute for Life and Environment (A\u2010LIFE) Vrije Universiteit Amsterdam  Amsterdam the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6977-6603","authenticated-orcid":false,"given":"J.","family":"Vreede","sequence":"additional","affiliation":[{"name":"Van't Hoff Institute for Molecular Sciences University of Amsterdam  Amsterdam the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0658-4775","authenticated-orcid":false,"given":"M. C. M.","family":"van Loosdrecht","sequence":"additional","affiliation":[{"name":"Department of Biotechnology Delft University of Technology  Delft the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3674-598X","authenticated-orcid":false,"given":"R.","family":"Kort","sequence":"additional","affiliation":[{"name":"Amsterdam Institute for Life and Environment (A\u2010LIFE) Vrije Universiteit Amsterdam  Amsterdam the Netherlands"},{"name":"ARTIS\u2010Micropia  Amsterdam the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,10]]},"reference":[{"key":"e_1_2_14_2_1","unstructured":"Abraham M. A.Alekseenko C.Bergh et\u00a0al.2023.\u201cGROMACS 2023 Manual.\u201dAccessed January 22 2026.https:\/\/zenodo.org\/records\/7588711."},{"key":"e_1_2_14_3_1","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.softx.2015.06.001","article-title":"GROMACS: High Performance Molecular Simulations Through Multi\u2010Level Parallelism From Laptops to Supercomputers","volume":"1","author":"Abraham M. J.","year":"2015","journal-title":"SoftwareX"},{"key":"e_1_2_14_4_1","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1038\/s41586-024-07487-w","article-title":"Accurate Structure Prediction of Biomolecular Interactions With AlphaFold 3","volume":"630","author":"Abramson J.","year":"2024","journal-title":"Nature"},{"key":"e_1_2_14_5_1","unstructured":"Adams A. N. D. L. E.Griffin J.Burnie et\u00a0al.2025.\u201cGram\u2010Positive Bacterial Cell Wall Components Inhibit Herpes Simplex Virus Infection.\u201d2025.11.02.686169. Accessed November 24 2025.https:\/\/doi.org\/10.1101\/2025.11.02.686169v1."},{"key":"e_1_2_14_6_1","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.00880\u201014"},{"key":"e_1_2_14_7_1","doi-asserted-by":"crossref","DOI":"10.1128\/jb.00417-24","article-title":"Recent Insights Into Wzy Polymerases and Lipopolysaccharide O\u2010Antigen Biosynthesis","volume":"207","author":"Ascari A.","year":"2025","journal-title":"Journal of Bacteriology"},{"key":"e_1_2_14_8_1","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1186\/s12934-021-01575-6","article-title":"Lyophilized Alginate\u2010Based Microspheres Containing \n                     Lactobacillus fermentum\n                   D12, an Exopolysaccharides Producer, Contribute to the strain's Functionality In\u00a0Vitro","volume":"20","author":"Butorac K.","year":"2021","journal-title":"Microbial Cell Factories"},{"key":"e_1_2_14_9_1","doi-asserted-by":"crossref","DOI":"10.1038\/s41522-026-00937-5","article-title":"Exopolysaccharides of \n                     Lactobacillus crispatus\n                   Mediate Key Balancing Interactions With the Vaginal Mucosa","volume":"12","author":"Croatti V.","year":"2026","journal-title":"npj Biofilms and Microbiomes"},{"key":"e_1_2_14_10_1","doi-asserted-by":"publisher","DOI":"10.1093\/jambio\/lxag218"},{"key":"e_1_2_14_11_1","doi-asserted-by":"crossref","first-page":"10879","DOI":"10.1038\/s41467-024-55233-7","article-title":"Lactobacillus crispatus\n                   S\u2010Layer Proteins Modulate Innate Immune Response and Inflammation in the Lower Female Reproductive Tract","volume":"15","author":"Decout A.","year":"2024","journal-title":"Nature Communications"},{"key":"e_1_2_14_12_1","doi-asserted-by":"crossref","DOI":"10.3390\/microorganisms7100444","article-title":"A Diverse Repertoire of Exopolysaccharide Biosynthesis Gene Clusters in Lactobacillus Revealed by Comparative Analysis in 106 Sequenced Genomes","volume":"7","author":"Deo D.","year":"2019","journal-title":"Microorganisms"},{"key":"e_1_2_14_13_1","doi-asserted-by":"crossref","DOI":"10.1186\/s12879-016-1513-3","article-title":"Molecular Assessment of Bacterial Vaginosis by Lactobacillus Abundance and Species Diversity","volume":"16","author":"Dols J. A. M.","year":"2016","journal-title":"BMC Infectious Diseases"},{"key":"e_1_2_14_14_1","doi-asserted-by":"crossref","first-page":"73","DOI":"10.3390\/microorganisms9010073","article-title":"Probiogenomics Analysis of 97 \n                     Lactobacillus crispatus\n                   Strains as a Tool for the Identification of Promising Next\u2010Generation Probiotics","volume":"9","author":"Fontana F.","year":"2021","journal-title":"Microorganisms"},{"key":"e_1_2_14_15_1","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1038\/s41564-022-01083-2","article-title":"Towards a Deeper Understanding of the Vaginal Microbiota","volume":"7","author":"France M.","year":"2022","journal-title":"Nature Microbiology"},{"key":"e_1_2_14_16_1","doi-asserted-by":"crossref","DOI":"10.3390\/ijms23105590","article-title":"Genetic Elements Orchestrating \n                     Lactobacillus crispatus\n                   Glycogen Metabolism in the Vagina","volume":"23","author":"Hertzberger R.","year":"2022","journal-title":"International Journal of Molecular Sciences"},{"key":"e_1_2_14_17_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00284\u2010025\u201004459\u20109"},{"key":"e_1_2_14_18_1","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1038\/nmeth.4067","article-title":"CHARMM36m: An Improved Force Field for Folded and Intrinsically Disordered Proteins","volume":"14","author":"Huang J.","year":"2017","journal-title":"Nature Methods"},{"key":"e_1_2_14_19_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0308526"},{"key":"e_1_2_14_20_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2017.01865\/full"},{"key":"e_1_2_14_21_1","doi-asserted-by":"crossref","first-page":"926","DOI":"10.1063\/1.445869","article-title":"Comparison of Simple Potential Functions for Simulating Liquid Water","volume":"79","author":"Jorgensen W. L.","year":"1983","journal-title":"Journal of Chemical Physics"},{"key":"e_1_2_14_22_1","doi-asserted-by":"crossref","first-page":"1893","DOI":"10.1021\/ci300604z","article-title":"Lessons Learned in Empirical Scoring With Smina From the CSAR 2011 Benchmarking Exercise","volume":"53","author":"Koes D. R.","year":"2013","journal-title":"Journal of Chemical Information and Modeling"},{"key":"e_1_2_14_23_1","doi-asserted-by":"crossref","first-page":"4121","DOI":"10.1074\/jbc.272.7.4121","article-title":"UDP\u2010Galactofuranose Precursor Required for Formation of the Lipopolysaccharide O Antigen of \n                     Klebsiella pneumoniae\n                   Serotype O1 Is Synthesized by the Product of the rfbDKPO1 Gene","volume":"272","author":"K\u00f6plin R.","year":"1997","journal-title":"Journal of Biological Chemistry"},{"key":"e_1_2_14_24_1","doi-asserted-by":"crossref","DOI":"10.1186\/1471-2105-10-168","article-title":"Fpocket: An Open Source Platform for Ligand Pocket Detection","volume":"10","author":"Le Guilloux V.","year":"2009","journal-title":"BMC Bioinformatics"},{"key":"e_1_2_14_25_1","first-page":"107","volume-title":"Galactose: Structure and Function in Biology and Medicine","author":"Marino C.","year":"2014"},{"key":"e_1_2_14_26_1","article-title":"Profiling of Vaginal \n                     Lactobacillus jensenii\n                   Isolated From Preterm and Full\u2010Term Pregnancies Reveals Strain\u2010Specific Factors Relating to Host Interaction","volume":"9","author":"Nori S. R. C.","year":"2023","journal-title":"Microbial Genomics"},{"key":"e_1_2_14_27_1","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1038\/s41522-025-00682-1","article-title":"Strain\u2010Level Variation Among Vaginal Lactobacillus\n                  c\n                  rispatus and \n                     Lactobacillus iners\n                   as Identified by Comparative Metagenomics","volume":"11","author":"Nori S. R. C.","year":"2025","journal-title":"NPJ Biofilms Microbiomes"},{"key":"e_1_2_14_28_1","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.mib.2021.01.007","article-title":"Modifications of Cell Wall Polymers in Gram\u2010Positive Bacteria by Multi\u2010Component Transmembrane Glycosylation Systems","volume":"60","author":"Rismondo J.","year":"2021","journal-title":"Current Opinion in Microbiology"},{"key":"e_1_2_14_29_1","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1007\/978-1-4939-9154-9_4","volume-title":"Bacterial Polysaccharides: Methods and Protocols","author":"Sadovskaya I.","year":"2019"},{"key":"e_1_2_14_30_1","doi-asserted-by":"crossref","first-page":"20798","DOI":"10.1038\/s41598-025-08321-7","article-title":"Comparative Genomic Analysis of Bacteriocin Genes in \n                     Lactobacillus crispatus\n                   Strains","volume":"15","author":"Santarelli G.","year":"2025","journal-title":"Scientific Reports"},{"key":"e_1_2_14_31_1","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1038\/nmeth.2089","article-title":"NIH Image to ImageJ: 25\u2009Years of Image Analysis","volume":"9","author":"Schneider C. A.","year":"2012","journal-title":"Nature Methods"},{"key":"e_1_2_14_32_1","doi-asserted-by":"crossref","DOI":"10.3390\/ijms21103465","article-title":"Galactofuranose\u2010Related Enzymes: Challenges and Hopes","volume":"21","author":"Seni\u010dar M.","year":"2020","journal-title":"International Journal of Molecular Sciences"},{"key":"e_1_2_14_33_1","doi-asserted-by":"crossref","DOI":"10.1371\/journal.ppat.1013346","article-title":"Through Thick and Thin: The Vaginal Microbiome as Both Occupant and Healer","volume":"21","author":"Sillen M.","year":"2025","journal-title":"PLoS Pathogens"},{"key":"e_1_2_14_34_1","doi-asserted-by":"crossref","first-page":"2233","DOI":"10.1128\/AEM.03566-12","article-title":"Genome Instability in \n                     Lactobacillus rhamnosus\n                   GG","volume":"79","author":"Sybesma W.","year":"2013","journal-title":"Applied and Environmental Microbiology"},{"key":"e_1_2_14_35_1","doi-asserted-by":"crossref","first-page":"1774","DOI":"10.1111\/1751-7915.14305","article-title":"The Core Genome Evolution of \n                     Lactobacillus crispatus\n                   as a Driving Force for Niche Competition in the Human Vaginal Tract","volume":"16","author":"Tarracchini C.","year":"2023","journal-title":"Microbial Biotechnology"},{"key":"e_1_2_14_36_1","doi-asserted-by":"crossref","first-page":"17612","DOI":"10.1074\/jbc.RA119.009957","article-title":"A Dual\u2010Chain Assembly Pathway Generates the High Structural Diversity of Cell\u2010Wall Polysaccharides in \n                     Lactococcus lactis","volume":"294","author":"Theodorou I.","year":"2019","journal-title":"Journal of Biological Chemistry"},{"key":"e_1_2_14_37_1","doi-asserted-by":"crossref","DOI":"10.1186\/s40168-019-0667-9","article-title":"Comparative Genomics of Human \n                     Lactobacillus crispatus\n                   Isolates Reveals Genes for Glycosylation and Glycogen Degradation: Implications for in Vivo Dominance of the Vaginal Microbiota","volume":"7","author":"Van der Veer C.","year":"2019","journal-title":"Microbiome"}],"container-title":["Molecular Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/mmi.70112","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1111\/mmi.70112","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/mmi.70112","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T04:47:46Z","timestamp":1789102066000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/mmi.70112"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,10]]},"references-count":36,"alternative-id":["10.1111\/mmi.70112"],"URL":"https:\/\/doi.org\/10.1111\/mmi.70112","archive":["Portico"],"relation":{},"ISSN":["0950-382X","1365-2958"],"issn-type":[{"value":"0950-382X","type":"print"},{"value":"1365-2958","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,10]]},"assertion":[{"value":"2026-06-02","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-08-31","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-10","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"mmi.70112"}}