{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,21]],"date-time":"2026-07-21T16:01:34Z","timestamp":1784649694922,"version":"3.55.0"},"reference-count":36,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,7,21]],"date-time":"2026-07-21T00:00:00Z","timestamp":1784592000000},"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>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      The multidrug-resistant pathogen\n                      <jats:italic>Clostridioides difficile<\/jats:italic>\n                      (\n                      <jats:italic>C. difficile<\/jats:italic>\n                      ) presents a persistent clinical threat. While Multidrug and Toxic Compound Extrusion (MATE) transporters are recognized as xenobiotic efflux pumps, their pleiotropic roles in pathogen physiology, particularly in stress adaptation and virulence regulation, remain largely unexplored. Understanding how\n                      <jats:italic>C. difficile<\/jats:italic>\n                      adapts and thrives in the face of host defenses and antimicrobial pressures, potentially influencing gut microbiome dynamics, is crucial for combating\n                      <jats:italic>C. difficile<\/jats:italic>\n                      infection.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      We functionally characterized the MATE transporter gene CD20030 (\n                      <jats:italic>mate<\/jats:italic>\n                      ) in\n                      <jats:italic>C. difficile<\/jats:italic>\n                      630. A markerless deletion mutant (\u0394\n                      <jats:italic>mate<\/jats:italic>\n                      ) and a complemented strain were constructed using a CRISPR-Cas9 system. Phenotypic assays determining antimicrobial susceptibility, oxidative stress tolerance, autolysis, and cytotoxicity were integrated with comparative proteomic profiling to assess the physiological changes.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The \u0394\n                      <jats:italic>mate<\/jats:italic>\n                      mutant demonstrated broad-spectrum hypersensitivity to antibiotics and hydrogen peroxide, indicating the involvement of this transporter in intrinsic resistance and oxidative defense. The mutant exhibited reduced autolysis; however, toxin production (\n                      <jats:italic>tcdA<\/jats:italic>\n                      and\n                      <jats:italic>tcdB<\/jats:italic>\n                      ) and cytotoxicity were significantly upregulated. In soft agar assays, the mutant showed expanded surface spreading. Proteomic data identified a &amp;gt;10,000-fold downregulation of flagellar structural proteins (FliC, FlgC) and a concurrent upregulation of the surface adhesin CwpV. This molecular evidence indicates a \u201cswimming-to-sliding transition\u201d driven by metabolic stress, rather than active swimming motility. These phenotypic and proteomic shifts present a resource reallocation strategy, where the bacterium sacrifices energy-consuming flagellar assembly to prioritize survival and virulence, potentially altering its interaction with the gut epithelial surface and resident microbiota.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      The MATE transporter (CD20030) operates as a pleiotropic regulatory hub and metabolic sentinel in\n                      <jats:italic>C. difficile<\/jats:italic>\n                      . Its absence induces metabolic reprogramming that orchestrates a motility-virulence trade-off, linking multidrug resistance directly to bacterial pathogenesis. These physiological adaptations likely dictate the pathogen\u2032s colonization and persistence strategies within the gut niche, potentially perturbing the host-microbiome equilibrium during infection.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fmicb.2026.1865850","type":"journal-article","created":{"date-parts":[[2026,7,21]],"date-time":"2026-07-21T15:04:31Z","timestamp":1784646271000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Pleiotropic roles of the MATE transporter CD20030 in Clostridioides difficile: linking multidrug resistance to oxidative stress defense and virulence regulation"],"prefix":"10.3389","volume":"17","author":[{"given":"Jie","family":"Deng","sequence":"first","affiliation":[{"name":"Key Laboratory of Endemic and Ethnic Diseases, Ministry of Education, Guizhou Medical University","place":["Guiyang, Guizhou, China"]},{"name":"Collaborative Innovation Center for Prevention and Control of Endemic and Ethnic Regional Diseases Co-constructed by the Province and Ministry","place":["Guiyang, Guizhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fa Hui","family":"Chen","sequence":"additional","affiliation":[{"name":"Key Laboratory of Endemic and Ethnic Diseases, Ministry of Education, Guizhou Medical University","place":["Guiyang, Guizhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wen Jing","family":"Wu","sequence":"additional","affiliation":[{"name":"Key Laboratory of Endemic and Ethnic Diseases, Ministry of Education, Guizhou Medical University","place":["Guiyang, Guizhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ting Yu","family":"Huang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Endemic and Ethnic Diseases, Ministry of Education, Guizhou Medical University","place":["Guiyang, Guizhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gu Zhen","family":"Cui","sequence":"additional","affiliation":[{"name":"Key Laboratory of Endemic and Ethnic Diseases, Ministry of Education, Guizhou Medical University","place":["Guiyang, Guizhou, China"]},{"name":"Laboratory of Microbiology and Parasitology of the Education Department of Guizhou, Guizhou Medical University","place":["Guiyang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Hong","sequence":"additional","affiliation":[{"name":"Key Laboratory of Endemic and Ethnic Diseases, Ministry of Education, Guizhou Medical University","place":["Guiyang, Guizhou, China"]},{"name":"School of Basic Medical Sciences, Guizhou Medical University","place":["Guiyang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,7,21]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"102350","DOI":"10.1016\/j.bbrep.2025.102350","article-title":"CdmR, a MarR family protein from Clostridioides difficile, binds with positive cooperativity in the predicted promoter region of a MATE efflux transporter","volume":"44","author":"Canepa","year":"2025","journal-title":"Biochem. Biophys. Rep."},{"key":"ref2001","doi-asserted-by":"publisher","first-page":"2358","DOI":"10.1016\/j.csbj.2024.05.027","article-title":"The complete catalog of antimicrobial resistance secondary active transporters in Clostridioides difficile: evolution and drug resistance perspective","volume":"23","author":"Chanket","year":"2024","journal-title":"Comput. Struct. Biotechnol. J."},{"key":"ref2","doi-asserted-by":"publisher","first-page":"1691769","DOI":"10.3389\/fmicb.2025.1691769","article-title":"Pig-L mediates virulence, biofilm formation, and oxidative stress tolerance in Clostridioides difficile","volume":"16","author":"Cheng","year":"2025","journal-title":"Front. Microbiol."},{"key":"ref3","doi-asserted-by":"publisher","first-page":"2343","DOI":"10.1099\/mic.0.27878-0","article-title":"Acd, a peptidoglycan hydrolase of Clostridium difficile with N-acetylglucosaminidase activity","volume":"151","author":"Dhalluin","year":"2005","journal-title":"Microbiology"},{"key":"ref4","doi-asserted-by":"publisher","first-page":"e00135-23","DOI":"10.1128\/cmr.00135-23","article-title":"Clostridioides difficile infection: history, epidemiology, risk factors, prevention, clinical manifestations, treatment, and future options","volume":"37","author":"Di Bella","year":"2024","journal-title":"Clin. Microbiol. Rev."},{"key":"ref5","doi-asserted-by":"publisher","first-page":"191","DOI":"10.1089\/mdr.2004.10.191","article-title":"CdeA of Clostridium difficile, a new multidrug efflux transporter of the MATE family","volume":"10","author":"Dridi","year":"2004","journal-title":"Microb. Drug Resist."},{"key":"ref6","doi-asserted-by":"publisher","first-page":"1698","DOI":"10.3389\/fmicb.2016.01698","article-title":"Chemical and stress resistances of Clostridium difficile spores and vegetative cells","volume":"7","author":"Edwards","year":"2016","journal-title":"Front. Microbiol."},{"key":"ref7","doi-asserted-by":"publisher","first-page":"402","DOI":"10.3390\/ph18030402","article-title":"Combating antibiotic resistance: mechanisms, multidrug-resistant pathogens, and novel therapeutic approaches: an updated review","volume":"18","author":"Elshobary","year":"2025","journal-title":"Pharmaceuticals (Basel)"},{"key":"ref8","doi-asserted-by":"publisher","first-page":"541","DOI":"10.1111\/j.1365-2958.2009.06812.x","article-title":"A novel genetic switch controls phase variable expression of CwpV, a Clostridium difficile cell wall protein","volume":"74","author":"Emerson","year":"2009","journal-title":"Mol. Microbiol."},{"key":"ref9","doi-asserted-by":"publisher","first-page":"e3000379","DOI":"10.1371\/journal.pbio.3000379","article-title":"Phase variation of a signal transduction system controls clostridioides difficile colony morphology, motility, and virulence","volume":"17","author":"Garrett","year":"2019","journal-title":"PLoS Biol."},{"key":"ref10","doi-asserted-by":"publisher","first-page":"e1000931","DOI":"10.1371\/journal.pgen.1000931","article-title":"A MATE-family efflux pump rescues the Escherichia coli 8-Oxoguanine-repair-deficient Mutator phenotype and protects against H2O2 killing","volume":"6","author":"Guelfo","year":"2010","journal-title":"PLoS Genet."},{"key":"ref11","doi-asserted-by":"publisher","first-page":"e106340","DOI":"10.1371\/journal.pone.0106340","article-title":"Overexpression of the novel MATE fluoroquinolone efflux pump FepA in Listeria monocytogenes is driven by inactivation of its local repressor FepR","volume":"9","author":"Gu\u00e9rin","year":"2014","journal-title":"PLoS One"},{"key":"ref12","doi-asserted-by":"publisher","first-page":"1588","DOI":"10.1021\/acssynbio.8b00087","article-title":"Multiplexed CRISPR-Cpf1-mediated genome editing in Clostridium difficile toward the understanding of pathogenesis of C. difficile infection","volume":"7","author":"Hong","year":"2018","journal-title":"ACS Synth. Biol."},{"key":"ref13","doi-asserted-by":"publisher","first-page":"1586662","DOI":"10.3389\/fmicb.2025.1586662","article-title":"Evaluation of protein extraction methodologies on bacterial proteomic profiling: a comparative analysis","volume":"16","author":"Jiang","year":"2025","journal-title":"Front. Microbiol."},{"key":"ref14","doi-asserted-by":"publisher","first-page":"796025","DOI":"10.3389\/fmicb.2021.796025","article-title":"The power of microbiome studies: some considerations on which alpha and Beta metrics to use and how to report results","volume":"12","author":"Kers","year":"2022","journal-title":"Front. Microbiol."},{"key":"ref15","doi-asserted-by":"publisher","first-page":"763","DOI":"10.1016\/j.bbapap.2008.11.012","article-title":"Multidrug efflux transporters in the MATE family","volume":"1794","author":"Kuroda","year":"2009","journal-title":"Biochim. Biophys. Acta"},{"key":"ref16","doi-asserted-by":"publisher","first-page":"183154","DOI":"10.1016\/j.bbamem.2019.183154","article-title":"Structural biology of the multidrug and toxic compound extrusion superfamily transporters","volume":"1862","author":"Kusakizako","year":"2020","journal-title":"Biochim. Biophys. Acta Biomembr."},{"key":"ref17","doi-asserted-by":"publisher","first-page":"823","DOI":"10.1128\/JB.02024-12","article-title":"Activity of Proteus mirabilis FliL is viscosity dependent and requires Extragenic DNA","volume":"195","author":"Lee","year":"2013","journal-title":"J. Bacteriol."},{"key":"ref18","doi-asserted-by":"publisher","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2\u2212\u0394\u0394CT method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref19","doi-asserted-by":"publisher","first-page":"e1013471","DOI":"10.1371\/journal.ppat.1013471","article-title":"Phase variation of clostridioides difficile colony morphology occurs via modulation of cell division","volume":"21","author":"Mehra","year":"2025","journal-title":"PLoS Pathog."},{"key":"ref20","doi-asserted-by":"publisher","first-page":"1610","DOI":"10.1128\/JB.01724-12","article-title":"Characterization of the MSMEG_2631 gene (mmp) encoding a multidrug and toxic compound extrusion (MATE) family protein in mycobacterium smegmatis and exploration of its Polyspecific nature using biolog phenotype MicroArray","volume":"195","author":"Mishra","year":"2013","journal-title":"J. Bacteriol."},{"key":"ref21","doi-asserted-by":"publisher","first-page":"587","DOI":"10.1016\/j.tips.2006.09.001","article-title":"The MATE proteins as fundamental transporters of metabolic and xenobiotic organic cations","volume":"27","author":"Omote","year":"2006","journal-title":"Trends Pharmacol. Sci."},{"key":"ref22","doi-asserted-by":"publisher","first-page":"e1002024","DOI":"10.1371\/journal.ppat.1002024","article-title":"The Clostridium difficile cell wall protein CwpV is antigenically variable between strains, but exhibits conserved aggregation-promoting function","volume":"7","author":"Reynolds","year":"2011","journal-title":"PLoS Pathog."},{"key":"ref23","doi-asserted-by":"publisher","first-page":"526","DOI":"10.1038\/nrmicro2164","article-title":"Clostridium difficile infection: new developments in epidemiology and pathogenesis","volume":"7","author":"Rupnik","year":"2009","journal-title":"Nat. Rev. Microbiol."},{"key":"ref24","doi-asserted-by":"publisher","first-page":"337","DOI":"10.1038\/nature10098","article-title":"Global quantification of mammalian gene expression control","volume":"473","author":"Schwanh\u00e4usser","year":"2011","journal-title":"Nature"},{"key":"ref25","doi-asserted-by":"publisher","first-page":"155","DOI":"10.1007\/5584_2023_771","article-title":"Different efflux pump systems in Acinetobacter baumannii and their role in multidrug resistance","volume":"1434","author":"Sharma","year":"2023","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref26","doi-asserted-by":"publisher","first-page":"15","DOI":"10.1007\/s00232-017-9992-8","article-title":"Biocide selective TolC-independent efflux pumps in Enterobacteriaceae","volume":"251","author":"Slipski","year":"2018","journal-title":"J. Membr. Biol."},{"key":"ref27","doi-asserted-by":"publisher","first-page":"227","DOI":"10.1038\/nrg3185","article-title":"Insights into the regulation of protein abundance from proteomic and transcriptomic analyses","volume":"13","author":"Vogel","year":"2012","journal-title":"Nat. Rev. Genet."},{"key":"ref28","doi-asserted-by":"publisher","first-page":"819","DOI":"10.1099\/00221287-136-5-819","article-title":"Conjugative plasmid transfer from Escherichia coli to Clostridium acetobutylicum","volume":"136","author":"Williams","year":"1990","journal-title":"J. Gen. Microbiol."},{"key":"ref29","doi-asserted-by":"publisher","first-page":"e00648-18","DOI":"10.1128\/mBio.00648-18","article-title":"Cwp19 is a novel lytic transglycosylase involved in stationary-phase autolysis resulting in toxin release in Clostridium difficile","volume":"9","author":"Wydau-Dematteis","year":"2018","journal-title":"MBio"},{"key":"ref30","doi-asserted-by":"publisher","first-page":"1028","DOI":"10.16288\/j.yczz.23-193","article-title":"Progress on mechanisms of antibiotic resistance in Clostridioides difficile","volume":"45","author":"Xu","year":"2023","journal-title":"Yi Chuan"},{"key":"ref31","doi-asserted-by":"publisher","first-page":"106570","DOI":"10.1016\/j.ijantimicag.2022.106570","article-title":"Metronidazole-resistant Clostridioides difficile: genomic and transcriptomic traits acquired under in-vitro metronidazole induction","volume":"59","author":"Xu","year":"2022","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref32","doi-asserted-by":"publisher","first-page":"1569627","DOI":"10.3389\/fmicb.2025.1569627","article-title":"The sigma factor \u03c354 (rpoN) functions as a global regulator of antibiotic resistance, motility, metabolism, and virulence in Clostridioides difficile","volume":"16","author":"Yang","year":"2025","journal-title":"Front. Microbiol."},{"key":"ref33","doi-asserted-by":"publisher","first-page":"2911","DOI":"10.1002\/bit.27435","article-title":"Enhancing plasmid transformation efficiency and enabling CRISPR-Cas9\/Cpf1-based genome editing in Clostridium tyrobutyricum","volume":"117","author":"Zhang","year":"2020","journal-title":"Biotechnol. Bioeng."},{"key":"ref34","doi-asserted-by":"publisher","first-page":"e0270421","DOI":"10.1128\/spectrum.02704-21","article-title":"The cwp66 gene affects cell adhesion, stress tolerance, and antibiotic resistance in Clostridioides difficile","volume":"10","author":"Zhou","year":"2022","journal-title":"Microbiol. Spectrum"},{"key":"ref35","doi-asserted-by":"publisher","first-page":"e03801","DOI":"10.1128\/mbio.03801-24","article-title":"Control of clostridioides difficile virulence and physiology by the flagellin homeostasis checkpoint FliC-FliW-CsrA in the absence of motility","volume":"16","author":"Zhu","year":"2025","journal-title":"MBio"}],"container-title":["Frontiers in Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2026.1865850\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,21]],"date-time":"2026-07-21T15:04:32Z","timestamp":1784646272000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2026.1865850\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,21]]},"references-count":36,"alternative-id":["10.3389\/fmicb.2026.1865850"],"URL":"https:\/\/doi.org\/10.3389\/fmicb.2026.1865850","relation":{},"ISSN":["1664-302X"],"issn-type":[{"value":"1664-302X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,21]]},"article-number":"1865850"}}