{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T17:53:23Z","timestamp":1791395603254,"version":"4.3.3"},"reference-count":30,"publisher":"Microbiology Society","issue":"10","license":[{"start":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T00:00:00Z","timestamp":1791331200000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Department for the Economy","id":[{"id":"https:\/\/ror.org\/0161w0r98","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,7]]},"abstract":"<jats:p>\n                    <jats:bold>Introduction.<\/jats:bold>\n                    <jats:italic>Clostridioides difficile<\/jats:italic>\n                    is an opportunistic enteric pathogen posing significant challenges in human and animal healthcare due to the recurrent nature of infections and associated financial burden. The molecular chaperone protein, DnaK (Hsp70), encoded by\n                    <jats:italic>dnaK<\/jats:italic>\n                    , plays a central role in the heat stress response and protein homeostasis. Disruption of\n                    <jats:italic>dnaK<\/jats:italic>\n                    in\n                    <jats:italic>C. difficile<\/jats:italic>\n                    630\u0394\n                    <jats:italic>erm<\/jats:italic>\n                    has previously been associated with increased biofilm formation and multiple phenotypic alterations.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Gap Statement.<\/jats:bold>\n                    Although disruption of\n                    <jats:italic>dnaK<\/jats:italic>\n                    has been shown to alter biofilm formation and multiple phenotypes in\n                    <jats:italic>C. difficile<\/jats:italic>\n                    , the global transcriptional mechanisms linking DnaK to biofilm development and stress adaptation remain poorly understood.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Aim.<\/jats:bold>\n                    This study aimed to characterize the global transcriptional response associated with\n                    <jats:italic>dnaK<\/jats:italic>\n                    disruption during planktonic and biofilm growth to identify pathways linking the DnaK-mediated stress response with biofilm formation.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Methods.<\/jats:bold>\n                    This study investigated the global transcriptional changes associated with\n                    <jats:italic>dnaK<\/jats:italic>\n                    disruption during planktonic and colony biofilm culture. Colony biofilms of\n                    <jats:italic>C. difficile<\/jats:italic>\n                    strains 630, 630\u0394\n                    <jats:italic>erm<\/jats:italic>\n                    and 630\u0394\n                    <jats:italic>erm:dnaK<\/jats:italic>\n                    were cultured on mixed nitrocellulose ester semi-permeable membranes and examined structurally using low-vacuum scanning electron microscopy. Total RNA was extracted from both planktonic cultures and 24 h colony biofilms, followed by Illumina RNA sequencing. Differential gene expression analysis was performed using DESeq (\n                    <jats:italic>P<\/jats:italic>\n                    <jats:sub>adj<\/jats:sub>\n                    &lt;0.05). Gene set enrichment analysis (Kyoto Encyclopedia of Genes and Genomes [KEGG] and Gene Ontology) was conducted to identify significantly enriched metabolic pathways and biological processes.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Results.<\/jats:bold>\n                    Disruption of\n                    <jats:italic>dnaK<\/jats:italic>\n                    resulted in global transcriptomic remodelling, particularly during biofilm culture, where 72.4% of genes were significantly differentially expressed relative to planktonic culture. A core number of pathways were involved in biofilm formation in all strains, including increased sporulation with reduced translation, amino acid biosynthesis and flagellar assembly. The\n                    <jats:italic>dnaK<\/jats:italic>\n                    mutant exhibited a unique pattern of enrichment of pathways associated with cell division, peptidoglycan synthesis, carbon metabolism and amino acid biosynthesis, alongside a coordinated reduction of motility and chemotaxis pathways.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Conclusion.<\/jats:bold>\n                    Disruption of the\n                    <jats:italic>dnaK<\/jats:italic>\n                    gene appears to link the stress response to biofilm formation through a number of pathways associated with biofilm development. Understanding this link may identify systems relevant to persistent and recurrent infections.\n                  <\/jats:p>","DOI":"10.1099\/jmm.0.002212","type":"journal-article","created":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T16:17:37Z","timestamp":1791389857000},"source":"Crossref","is-referenced-by-count":0,"title":["Disruption of dnaK results in transcriptional reprogramming of biofilm-associated pathways in Clostridioides difficile"],"prefix":"10.1099","volume":"75","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2565-2705","authenticated-orcid":false,"given":"Megan G.","family":"Taggart","sequence":"first","affiliation":[{"name":"Nutrition Innovation Centre for Food & Health (NICHE), Ulster University, Coleraine, BT52 1SA, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5409-7512","authenticated-orcid":false,"given":"Carla S.","family":"M\u00f6ller-Levet","sequence":"additional","affiliation":[{"name":"Faculty of Health and Medical Sciences, University of Surrey, Guildford, Surrey GU2 7XH, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8204-1743","authenticated-orcid":false,"given":"William J.","family":"Snelling","sequence":"additional","affiliation":[{"name":"Nutrition Innovation Centre for Food & Health (NICHE), Ulster University, Coleraine, BT52 1SA, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5995-9468","authenticated-orcid":false,"given":"Patrick J.","family":"Naughton","sequence":"additional","affiliation":[{"name":"Nutrition Innovation Centre for Food & Health (NICHE), Ulster University, Coleraine, BT52 1SA, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5861-613X","authenticated-orcid":false,"given":"Roberto M.","family":"La Ragione","sequence":"additional","affiliation":[{"name":"School of Veterinary Medicine, University of Surrey, Guildford, Surrey GU2 7AL, UK"},{"name":"School of Biosciences, University of Surrey, Guildford, Surrey GU2 7XH, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barry M. G.","family":"O'Hagan","sequence":"additional","affiliation":[{"name":"Genomic Medicine Research Group, Ulster University, Coleraine, BT52 1SA, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9459-5572","authenticated-orcid":false,"given":"James S. G.","family":"Dooley","sequence":"additional","affiliation":[{"name":"Nutrition Innovation Centre for Food & Health (NICHE), Ulster University, Coleraine, BT52 1SA, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1788-7571","authenticated-orcid":false,"given":"Nigel G.","family":"Ternan","sequence":"additional","affiliation":[{"name":"Nutrition Innovation Centre for Food & Health (NICHE), Ulster University, Coleraine, BT52 1SA, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"345","reference":[{"key":"R1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1009348"},{"key":"R2","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms11102525"},{"key":"R3","doi-asserted-by":"publisher","DOI":"10.1016\/j.anaerobe.2025.102965"},{"key":"R4","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa0907635"},{"key":"R5","doi-asserted-by":"publisher","DOI":"10.1016\/j.anaerobe.2019.102108"},{"key":"R6","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-025-64067-w"},{"key":"R7","doi-asserted-by":"publisher","DOI":"10.1038\/s41522-021-00184-w"},{"key":"R8","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0050527"},{"key":"R9","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00147-12"},{"key":"R10","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pgen.1003660"},{"key":"R11","doi-asserted-by":"publisher","DOI":"10.1111\/j.1469-0691.2012.03853.x"},{"key":"R12","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1009817"},{"key":"R13","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01980-12"},{"key":"R14","doi-asserted-by":"publisher","DOI":"10.1021\/pr200327t"},{"key":"R15","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0042410"},{"key":"R16","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0088960"},{"key":"R17","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-17583-9"},{"key":"R18","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkx353"},{"key":"R19","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/29.9.e45"},{"key":"R20","volume-title":"The Role of Clostridium Difficile Surface Structures in Virulence and Potential for Host Infection","author":"Alotaibi","year":"2017"},{"key":"R21","article-title":"Investigation of the Factors Regulating Biofilm Formation in Antibiotic Resistant Clostridioides Difficile Using Transcriptomics","author":"Taggart","year":"2023"},{"key":"R22","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2018.01970"},{"key":"R23","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00035-10"},{"key":"R24","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-35050-x"},{"key":"R25","doi-asserted-by":"publisher","DOI":"10.1093\/femspd\/ftw061"},{"key":"R26","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2018.02084"},{"key":"R27","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M116.749523"},{"key":"R28","doi-asserted-by":"publisher","DOI":"10.1099\/jmm.0.45790-0"},{"key":"R29","doi-asserted-by":"publisher","DOI":"10.1186\/s12864-015-1252-7"},{"key":"R30","doi-asserted-by":"publisher","DOI":"10.1038\/ng1830"}],"container-title":["Journal of Medical Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jmm\/10.1099\/jmm.0.002212?crawler=true&mimetype=application\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T16:17:41Z","timestamp":1791389861000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jmm\/10.1099\/jmm.0.002212"}},"subtitle":["This article is part of the Clostridioides difficile collection."],"short-title":[],"issued":{"date-parts":[[2026,10,7]]},"references-count":30,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2026,10,7]]}},"URL":"https:\/\/doi.org\/10.1099\/jmm.0.002212","relation":{},"ISSN":["0022-2615","1473-5644"],"issn-type":[{"value":"0022-2615","type":"print"},{"value":"1473-5644","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10,7]]}}}