{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T12:07:41Z","timestamp":1788782861086,"version":"build-2803163510"},"reference-count":63,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T00:00:00Z","timestamp":1788739200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T00:00:00Z","timestamp":1788739200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["MicrobiologyOpen"],"published-print":{"date-parts":[[2026,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Pyrimidine biosynthesis is essential for bacterial growth, but its role in regulating biofilm formation in\n                    <jats:italic>Francisella (F.) novicida<\/jats:italic>\n                    remains poorly defined. In this study, we experimentally defined the\n                    <jats:italic>carABpyrB<\/jats:italic>\n                    operon in\n                    <jats:italic>F. novicida<\/jats:italic>\n                    and investigated how disruption of the de novo pyrimidine biosynthesis pathway affects growth and biofilm formation under nutrient\u2010restricted conditions. Reverse transcriptase PCR confirmed co\u2010transcription of\n                    <jats:italic>carA<\/jats:italic>\n                    ,\n                    <jats:italic>carB<\/jats:italic>\n                    , and\n                    <jats:italic>pyrB<\/jats:italic>\n                    , and promoter prediction identified two putative \u03c3\n                    <jats:sup>70<\/jats:sup>\n                    \u2010dependent promoter regions upstream of\n                    <jats:italic>carA<\/jats:italic>\n                    . Transposon mutants disrupted in\n                    <jats:italic>carA<\/jats:italic>\n                    ,\n                    <jats:italic>carB<\/jats:italic>\n                    , and\n                    <jats:italic>pyrB<\/jats:italic>\n                    exhibited pronounced growth defects in Chamberlain's Defined Medium that were restored by uracil supplementation, confirming pyrimidine auxotrophy and functional disruption of de novo pyrimidine biosynthesis. We then extended this analysis to additional genes in the pyrimidine biosynthetic pathway and assessed biofilm formation in modified Mueller\u2013Hinton broth, a nutrient\u2010restricted condition. In this medium,\n                    <jats:italic>carA<\/jats:italic>\n                    ,\n                    <jats:italic>carB<\/jats:italic>\n                    ,\n                    <jats:italic>pyrB<\/jats:italic>\n                    , and\n                    <jats:italic>pyrE<\/jats:italic>\n                    mutants exhibited growth deficiencies; however, the\n                    <jats:italic>pyrE<\/jats:italic>\n                    mutant uniquely produced significantly more biofilm than the wild type. This phenotype remained evident even without growth normalization, with the\n                    <jats:italic>pyrE<\/jats:italic>\n                    mutant producing 3.3\u2010fold more biofilm than wild type, despite impaired growth, and increased to 11.8\u2010fold when normalized to growth. Quantitative PCR demonstrated that uracil supplementation represses\n                    <jats:italic>carA<\/jats:italic>\n                    ,\n                    <jats:italic>carB<\/jats:italic>\n                    , and\n                    <jats:italic>pyrB<\/jats:italic>\n                    transcription, consistent with feedback regulation of the pathway. Together, these findings indicate that pyrimidine limitation is not simply a growth\u2010limiting condition but can alter biofilm regulation, with\n                    <jats:italic>pyrE<\/jats:italic>\n                    disruption revealing a strong association between de novo pyrimidine biosynthesis and biofilm formation.\n                  <\/jats:p>","DOI":"10.1002\/mbo3.70355","type":"journal-article","created":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T11:10:04Z","timestamp":1788779404000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Characterization of\n                    <i>carABpyrB<\/i>\n                    operon and role of\n                    <i>pyrE<\/i>\n                    in\n                    <i>Francisella novicida<\/i>\n                    biofilm"],"prefix":"10.1002","volume":"15","author":[{"given":"Katie C.","family":"Sprinkel","sequence":"first","affiliation":[{"name":"School of Systems Biology George Mason University Manassas Virginia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Annika R.","family":"Geiger","sequence":"additional","affiliation":[{"name":"School of Systems Biology George Mason University Manassas Virginia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Monique L.","family":"van Hoek","sequence":"additional","affiliation":[{"name":"School of Systems Biology George Mason University Manassas Virginia USA"},{"name":"Center for Infectious Disease Research George Mason University Manassas Virginia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,7]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00146\u201010"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.00575\u201018"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462\u20102920.2010.02229.x"},{"key":"e_1_2_12_5_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0077834"},{"key":"e_1_2_12_6_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2020.593542"},{"key":"e_1_2_12_7_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsinfecdis.9b00411"},{"key":"e_1_2_12_8_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.81.13.4139"},{"key":"e_1_2_12_9_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12866\u2010016\u20100819\u2010z"},{"key":"e_1_2_12_10_1","doi-asserted-by":"publisher","DOI":"10.1128\/am.13.2.232\u2010235.1965"},{"key":"e_1_2_12_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598\u2010019\u201048697\u2010x"},{"key":"e_1_2_12_12_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00726\u2010018\u20102654\u2010z"},{"key":"e_1_2_12_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/0022\u20102836(88)90047\u20102"},{"key":"e_1_2_12_14_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0093119"},{"key":"e_1_2_12_15_1","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.031021\u20100"},{"key":"e_1_2_12_16_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.02165\u201015"},{"key":"e_1_2_12_17_1","doi-asserted-by":"publisher","DOI":"10.1080\/21505594.2015.1046029"},{"key":"e_1_2_12_18_1","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2020.00082"},{"key":"e_1_2_12_19_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.00115\u201025"},{"key":"e_1_2_12_20_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00248\u2010009\u20109586\u20109"},{"key":"e_1_2_12_21_1","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00001\u201012"},{"key":"e_1_2_12_22_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0606713104"},{"issue":"10","key":"e_1_2_12_23_1","doi-asserted-by":"crossref","first-page":"4479","DOI":"10.1128\/IAI.00610-08","article-title":"Adaptation of Francisella Tularensis to the Mammalian Environment is Governed by Cues Which Can be Mimicked In Vitro","volume":"76","author":"Hazlett K. 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