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The molecular mechanisms of GI colonization by\n                      <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                      include potential interactions with host cells and the mucus layer formed from secreted mucin glycoproteins.\n                      <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                      associates with the mucus layer\n                      <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                      and will associate with both epithelial cells and mucosal surfaces\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      . Previously, we found a substantial defect in binding to mucosal surfaces for mutants of the major flagellar subunit,\n                      <jats:italic toggle=\"yes\">fliC<\/jats:italic>\n                      , while mutation of the major subunit of type IV pili,\n                      <jats:italic toggle=\"yes\">pilA1<\/jats:italic>\n                      , showed increased adhesion. To elucidate the mechanisms by which\n                      <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                      interacts with\n                      <jats:italic toggle=\"yes\">ex vivo<\/jats:italic>\n                      mucosal surfaces, we have measured swimming motility, mucosal adhesion, and levels of flagellation by transmission electron microscopy for mutants of flagellar and T4P genes in\n                      <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                      R20291. We discovered that the\n                      <jats:italic toggle=\"yes\">pilA1<\/jats:italic>\n                      mutant showed increased flagellation, while decreases in flagellation were found for\n                      <jats:italic toggle=\"yes\">fliC, fliD,<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">flg<\/jats:italic>\n                      -\u03943 OFF (a phase-locked mutant with low transcription of the F3 flagellar operon), which were associated with both low swimming motility and low adhesion to mucosal surfaces. However, the reversed\n                      <jats:italic toggle=\"yes\">flg<\/jats:italic>\n                      -\u03943 ON mutant showed increased flagellation without a significant increase in adhesion. We also found that the\n                      <jats:italic toggle=\"yes\">fliC<\/jats:italic>\n                      mutant was defective in binding to mucus-secreting HT-29 MTX cells, and that decreased binding was not observed for other mutants with reduced flagellation. These results imply that at least two molecular pathways contribute to\n                      <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                      mucosal adhesion. In addition to their direct roles encoding T4P and flagellar subunits,\n                      <jats:italic toggle=\"yes\">pilA1<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">fliC<\/jats:italic>\n                      may contribute to regulatory networks governing other proteins relevant to mucosal adhesion.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        In the context of previous work on\n                        <jats:italic toggle=\"yes\">Clostridioides difficile<\/jats:italic>\n                        host adhesion by our groups and others, our results suggest that (i) at least two mechanisms exist for mucosal adhesion by\n                        <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                        , potentially direct adhesion by flagella and another lectin-like adhesion regulated by flagellar components; (ii) mucosal binding can also contribute to\n                        <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                        adhesion in 2D cell culture and could explain previous defects from\n                        <jats:italic toggle=\"yes\">fliC<\/jats:italic>\n                        mutants; and (iii) levels of flagellation are largely insensitive to\n                        <jats:italic toggle=\"yes\">fliC<\/jats:italic>\n                        transcription, implying that other factors limit flagellar production and that FliC levels may be regulated independently as part of regulatory networks within\n                        <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                        .\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/jb.00428-25","type":"journal-article","created":{"date-parts":[[2025,11,24]],"date-time":"2025-11-24T14:01:13Z","timestamp":1763992873000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Dissecting the role of flagellar subunits in\n                    <i>C. difficile<\/i>\n                    mucosal colonization"],"prefix":"10.1128","volume":"207","author":[{"given":"Baishakhi","family":"Biswas","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/043mer456","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Food Science and Technology, University of Nebraska-Lincoln","place":["Lincoln, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thi Van Thanh","family":"Do","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/043mer456","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Food Science and Technology, University of Nebraska-Lincoln","place":["Lincoln, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Justin G.","family":"Perdomo","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/043mer456","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biochemistry, University of Nebraska-Lincoln","place":["Lincoln, USA"]},{"id":[{"id":"https:\/\/ror.org\/043mer456","id-type":"ROR","asserted-by":"publisher"}],"name":"Nebraska Food for Health Center, University of Nebraska-Lincoln","place":["Lincoln, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3038-583X","authenticated-orcid":true,"given":"Jennifer M.","family":"Auchtung","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/043mer456","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Food Science and Technology, University of Nebraska-Lincoln","place":["Lincoln, USA"]},{"id":[{"id":"https:\/\/ror.org\/043mer456","id-type":"ROR","asserted-by":"publisher"}],"name":"Nebraska Food for Health Center, University of Nebraska-Lincoln","place":["Lincoln, USA"]},{"id":[{"id":"https:\/\/ror.org\/043mer456","id-type":"ROR","asserted-by":"publisher"}],"name":"Center for Integrated Biomolecular Communication, University of Nebraska-Lincoln","place":["Lincoln, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2527-0301","authenticated-orcid":true,"given":"Kurt H.","family":"Piepenbrink","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/043mer456","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Food Science and Technology, University of Nebraska-Lincoln","place":["Lincoln, USA"]},{"id":[{"id":"https:\/\/ror.org\/043mer456","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biochemistry, University of Nebraska-Lincoln","place":["Lincoln, USA"]},{"id":[{"id":"https:\/\/ror.org\/043mer456","id-type":"ROR","asserted-by":"publisher"}],"name":"Nebraska Food for Health Center, University of Nebraska-Lincoln","place":["Lincoln, USA"]},{"id":[{"id":"https:\/\/ror.org\/043mer456","id-type":"ROR","asserted-by":"publisher"}],"name":"Center for Integrated Biomolecular Communication, University of Nebraska-Lincoln","place":["Lincoln, USA"]},{"id":[{"id":"https:\/\/ror.org\/043mer456","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, University of Nebraska-Lincoln","place":["Lincoln, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2020.11.034"},{"key":"e_1_3_3_3_2","volume-title":"Antibiotic resistance threats in the United States","author":"CDC","year":"2019","unstructured":"CDC. 2019. 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