{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T16:57:44Z","timestamp":1785430664928,"version":"3.56.0"},"posted":{"date-parts":[[2026,5,6]]},"group-title":"Microbiology","reference-count":37,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,5,6]],"date-time":"2026-05-06T00:00:00Z","timestamp":1778025600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"award":["R01 AI132606"],"award-info":[{"award-number":["R01 AI132606"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01 AI153521"],"award-info":[{"award-number":["R01 AI153521"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  <jats:italic>Neisseria gonorrhoeae<\/jats:italic>\n                  is a major public health concern due to its high global prevalence and rapid evolution of antibiotic resistance. A first-in-class topoisomerase inhibitor, zoliflodacin (a spiropyrimidinetrione) recently received FDA approval for treatment of gonorrhea, but its potential for cross-resistance with another topoisomerase inhibitor, the fluoroquinolone antibiotic ciprofloxacin, remains poorly understood. Here, we investigated how genetic diversity in the fluoroquinolone target\n                  <jats:italic>gyrA<\/jats:italic>\n                  influences the resistance and fitness effects of the zoliflodacin resistance mutation\n                  <jats:italic>gyrB<\/jats:italic>\n                  <jats:sup>D429N<\/jats:sup>\n                  . We constructed an isogenic panel of\n                  <jats:italic>N. gonorrhoeae<\/jats:italic>\n                  to determine how the resistance and fitness effects of the\n                  <jats:italic>gyrB<\/jats:italic>\n                  <jats:sup>D429N<\/jats:sup>\n                  mutation are modulated by the most common ciprofloxacin resistance-associated variants in\n                  <jats:italic>gyrA<\/jats:italic>\n                  . In the presence of\n                  <jats:italic>gyrB<\/jats:italic>\n                  <jats:sup>D429N<\/jats:sup>\n                  , the zoliflodacin minimum inhibitory concentration (MIC) was 2-4-fold higher in strains that also contained ciprofloxacin resistance-associated\n                  <jats:italic>gyrA<\/jats:italic>\n                  alleles, and the\n                  <jats:italic>gyrB<\/jats:italic>\n                  <jats:sup>D429N<\/jats:sup>\n                  mutation reciprocally increased ciprofloxacin MICs of these strains 3-6-fold. Fitness cost of the\n                  <jats:italic>gyrB<\/jats:italic>\n                  <jats:sup>D429N<\/jats:sup>\n                  mutation varied from modest to severe across\n                  <jats:italic>gyrA<\/jats:italic>\n                  backgrounds, with the largest cost in ciprofloxacin resistant\n                  <jats:italic>gyrA<\/jats:italic>\n                  <jats:sup>91F\/95G<\/jats:sup>\n                  and\n                  <jats:italic>gyrA<\/jats:italic>\n                  <jats:sup>91F\/95N<\/jats:sup>\n                  backgrounds and comparatively minimal cost in the ciprofloxacin resistant\n                  <jats:italic>gyrA<\/jats:italic>\n                  <jats:sup>91F\/95A<\/jats:sup>\n                  background. These results demonstrate the capacity for epistatic interactions among resistance-associated\n                  <jats:italic>gyrA<\/jats:italic>\n                  and\n                  <jats:italic>gyrB<\/jats:italic>\n                  mutations, underscoring the need for genomic surveillance to monitor high-risk combinations of resistance determinants as new therapies are deployed.\n                <\/jats:p>","DOI":"10.64898\/2026.05.04.722797","type":"posted-content","created":{"date-parts":[[2026,5,7]],"date-time":"2026-05-07T07:50:14Z","timestamp":1778140214000},"source":"Crossref","is-referenced-by-count":1,"title":["Fluoroquinolone resistance-conferring\n                  <i>gyrA<\/i>\n                  variants alter the fitness cost and potentiate the resistance of the zoliflodacin resistance mutation\n                  <i>gyrB<\/i>\n                  <sup>D429N<\/sup>\n                  in\n                  <i>Neisseria gonorrhoeae<\/i>"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2208-9945","authenticated-orcid":false,"given":"Aditi","family":"Mukherjee","sequence":"first","affiliation":[{"name":"Department of Immunology and Infectious Diseases, Harvard T. H. Chan School of Public Health, Boston, Massachusetts, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8967-3564","authenticated-orcid":false,"given":"Sofia OP","family":"Blomqvist","sequence":"additional","affiliation":[{"name":"Department of Immunology and Infectious Diseases, Harvard T. H. Chan School of Public Health, Boston, Massachusetts, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9319-7097","authenticated-orcid":false,"given":"David","family":"Helekal","sequence":"additional","affiliation":[{"name":"Department of Immunology and Infectious Diseases, Harvard T. H. Chan School of Public Health, Boston, Massachusetts, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6753-4213","authenticated-orcid":false,"given":"Daniel HF","family":"Rubin","sequence":"additional","affiliation":[{"name":"Department of Immunology and Infectious Diseases, Harvard T. H. Chan School of Public Health, Boston, Massachusetts, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5908-9765","authenticated-orcid":false,"given":"Bailey","family":"Bowcutt","sequence":"additional","affiliation":[{"name":"Department of Immunology and Infectious Diseases, Harvard T. H. Chan School of Public Health, Boston, Massachusetts, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7849-8078","authenticated-orcid":false,"given":"Samantha G","family":"Palace","sequence":"additional","affiliation":[{"name":"Department of Immunology and Infectious Diseases, Harvard T. H. Chan School of Public Health, Boston, Massachusetts, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5646-1314","authenticated-orcid":false,"given":"Yonatan H","family":"Grad","sequence":"additional","affiliation":[{"name":"Department of Immunology and Infectious Diseases, Harvard T. H. Chan School of Public Health, Boston, Massachusetts, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026051105050566000_2026.05.04.722797v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(25)00118-5"},{"key":"2026051105050566000_2026.05.04.722797v1.2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00010-14"},{"key":"2026051105050566000_2026.05.04.722797v1.3","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/ciac043"},{"key":"2026051105050566000_2026.05.04.722797v1.4","unstructured":"U.S. Food and Drug Administration. FDA Approves Two Oral Therapies to Treat Gonorrhea 2025 [9 Mar 2026]. Available from: https:\/\/www.fda.gov\/news-events\/press-announcements\/fda-approves-two-oral-therapies-treat-gonorrhea."},{"key":"2026051105050566000_2026.05.04.722797v1.5","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.1994.tb01297.x"},{"key":"2026051105050566000_2026.05.04.722797v1.6","doi-asserted-by":"publisher","DOI":"10.15585\/mmwr.mm7312a2"},{"key":"2026051105050566000_2026.05.04.722797v1.7","unstructured":"Centers for Disease Control and Prevention. Sexually Transmitted Disease Surveillance January-July 2024: Gonococcal Isolate Surveillance Project National Profile. 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