{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,8,3]],"date-time":"2026-08-03T20:50:10Z","timestamp":1785790210530,"version":"3.56.0"},"posted":{"date-parts":[[2026,7,31]]},"group-title":"Microbiology","reference-count":60,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T00:00:00Z","timestamp":1785456000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["1R35GM162142-01"],"award-info":[{"award-number":["1R35GM162142-01"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["1R01AI198349-01"],"award-info":[{"award-number":["1R01AI198349-01"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,7,31]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Children with cystic fibrosis (CF) commonly harbor\n                  <jats:italic>Staphylococcus<\/jats:italic>\n                  aureus thymidine-dependent small-colony variants (TD-SCVs), which are associated with reduced lung function and increased respiratory exacerbations. How TD-SCVs survive in the thymidine-limited CF lung is unknown. Here, we show that TD-SCVs exhibit impaired glutamine uptake and depend on c-di-AMP-regulated\n                  <jats:italic>de novo<\/jats:italic>\n                  glutamine synthesis for survival in the murine lung. We found that transcription of the glutamine synthetase gene\n                  <jats:italic>glnA<\/jats:italic>\n                  is cooperatively repressed by the transcriptional regulator GlnR, the c-di-AMP-binding protein PstA, and GlnA itself. Glutamine starvation elevates c-di-AMP levels, relieving repression by this ternary complex and promoting glutamine synthesis. Reducing c-di-AMP levels causes a profound growth defect in TD-SCVs under low-thymidine conditions, which is rescued by\n                  <jats:italic>glnA<\/jats:italic>\n                  overexpression. Moreover, pharmacological inhibition of GlnA markedly impairs TD-SCV growth in murine lung. These findings elucidate the molecular mechanism underlying\n                  <jats:italic>S. aureus<\/jats:italic>\n                  TD-SCV survival during infection and identify glutamine synthesis as a promising therapeutic target for treating infections caused by antifolate-resistant bacteria.\n                <\/jats:p>","DOI":"10.64898\/2026.07.30.741254","type":"posted-content","created":{"date-parts":[[2026,8,1]],"date-time":"2026-08-01T04:50:19Z","timestamp":1785559819000},"source":"Crossref","is-referenced-by-count":0,"title":["A c-di-AMP-controlled glutamine synthesis pathway promotes persistence of\n                  <i>Staphylococcus aureus<\/i>\n                  thymidine-dependent small colony variants in the lung"],"prefix":"10.64898","author":[{"given":"Joshua P.","family":"Leeming","sequence":"first","affiliation":[{"name":"Department of Biology, University of Texas at Arlington, Arlington, TX, 76019, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shoukai","family":"Kang","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Washington, Seattle, WA 98195, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pearl P.","family":"Thakkar","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Texas at Arlington, Arlington, TX, 76019, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pratyusha","family":"Gorige","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Texas at Arlington, Arlington, TX, 76019, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Devanshi R.","family":"Patel","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Texas at Arlington, Arlington, TX, 76019, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuxin","family":"Pan","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Washington, Seattle, WA 98195, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Oscar E.","family":"Villasana Espinosa","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Texas at Arlington, Arlington, TX, 76019, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel J.","family":"Wolter","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, University of Washington, Seattle, WA, 98105, USA"},{"name":"Pulmonary and Sleep Medicine, Seattle Children\u2019s Hospital, Seattle, WA, 98105, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cara C.","family":"Boutte","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Texas at Arlington, Arlington, TX, 76019, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qing","family":"Tang","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Texas at Arlington, Arlington, TX, 76019, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026080313001369000_2026.07.30.741254v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(22)02185-7"},{"key":"2026080313001369000_2026.07.30.741254v1.2","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-020-03540-9"},{"key":"2026080313001369000_2026.07.30.741254v1.3","doi-asserted-by":"publisher","DOI":"10.1186\/s12890-015-0062-7"},{"key":"2026080313001369000_2026.07.30.741254v1.4","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens10091177"},{"key":"2026080313001369000_2026.07.30.741254v1.5","unstructured":"Foundation, C. 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