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The clinical utility of MLS antibiotics has diminished due to the dissemination of\n                      <jats:underline>e<\/jats:underline>\n                      rythromycin\n                      <jats:underline>r<\/jats:underline>\n                      esistance rRNA\n                      <jats:underline>m<\/jats:underline>\n                      ethyltransferase (Erm) genes.\n                      <jats:italic toggle=\"yes\">Staphylococcus aureus<\/jats:italic>\n                      ErmB methylates the universally conserved A2058 nucleotide of the 23S rRNA (m\n                      <jats:sup>6<\/jats:sup>\n                      A2058), resulting in cross-resistance to all MLS antibiotics by reducing drug-binding affinity. The operonic upstream ribosome stalling peptide ErmBL was thought to be the sole regulatory element controlling ErmB synthesis and the extent of MLS resistance. Unexpectedly, our laboratory evolution experiments revealed that numerous loss-of-function mutations outside the bicistronic\n                      <jats:italic toggle=\"yes\">ermBL-ermB<\/jats:italic>\n                      operon amplify ErmB-mediated MLS resistance. Among these are mutations in genes critical for purine\n                      <jats:italic toggle=\"yes\">de novo<\/jats:italic>\n                      biosynthesis and the salvage pathway. Specifically, inactivation of the gene encoding the purine biosynthesis transcriptional repressor (PurR) converts an otherwise moderately resistant\n                      <jats:italic toggle=\"yes\">\n                        ermBL-ermB (ermB\n                        <jats:sup>+<\/jats:sup>\n                      <\/jats:italic>\n                      ) strain into one exhibiting pronounced hyper-resistance to MLS antibiotics. This cooperative resistance phenotype is not attributable to increased\n                      <jats:italic toggle=\"yes\">ermB<\/jats:italic>\n                      expression or elevated m\n                      <jats:sup>6<\/jats:sup>\n                      A2058 modification. Instead,\n                      <jats:italic toggle=\"yes\">purR<\/jats:italic>\n                      inactivation leads to derepression of purine and pyrimidine biosynthesis, accompanied by increased expression of ribosome components. We find that the elevated ribosome abundance and translational capacity of the\n                      <jats:italic toggle=\"yes\">\n                        ermB\n                        <jats:sup>+<\/jats:sup>\n                      <\/jats:italic>\n                      \u2206\n                      <jats:italic toggle=\"yes\">purR<\/jats:italic>\n                      are directly proportional to its accelerated growth rate, thereby priming\n                      <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                      for survival under high MLS concentrations. These findings support a model in which expanded nucleotide metabolites and a surplus of antibiotic-free ribosomes drive global translation to buffer the inhibitory effects of MLS antibiotics.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        The Erm rRNA methyltransferase superfamily represents the most prevalent determinant of MLS resistance in nosocomial Gram-negative and Gram-positive bacteria. Previous studies have established that\n                        <jats:italic toggle=\"yes\">erm<\/jats:italic>\n                        expression is primarily governed by upstream ribosome stalling peptide and the 5\u2032 untranslated regions. Using the widespread\n                        <jats:italic toggle=\"yes\">S. aureus ermBL-ermB<\/jats:italic>\n                        (\n                        <jats:italic toggle=\"yes\">ermB<\/jats:italic>\n                        <jats:sup>+<\/jats:sup>\n                        ) operon as a model system, we unexpectedly identified second-site mutations in\n                        <jats:italic toggle=\"yes\">purR<\/jats:italic>\n                        that synergistically enhance MLS resistance in an\n                        <jats:italic toggle=\"yes\">ermB<\/jats:italic>\n                        <jats:sup>+<\/jats:sup>\n                        background. Loss of\n                        <jats:italic toggle=\"yes\">purR<\/jats:italic>\n                        function derepresses nucleotide biosynthesis and ribosome production, thereby promoting bacterial growth under antibiotic stress. While numerous\n                        <jats:italic toggle=\"yes\">purR<\/jats:italic>\n                        single-nucleotide polymorphisms across multiple species have been associated with antibiotic resistance, no study has directly linked these sequence polymorphisms to their regulatory function. Our results highlight the critical role of ribosome abundance and nucleotide metabolism in shaping antibiotic efficacy.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/mbio.00118-26","type":"journal-article","created":{"date-parts":[[2026,4,13]],"date-time":"2026-04-13T13:01:13Z","timestamp":1776085273000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Inactivation of a purine biosynthesis repressor promotes ribosome synthesis to overcome antibiotic stress"],"prefix":"10.1128","volume":"17","author":[{"given":"Alexandre","family":"Le Scornet","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02ets8c94","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology-Immunology, Northwestern University Feinberg School of Medicine","place":["Chicago, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongjun","family":"Tan","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01p7jjy08","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biology, College of Arts and Sciences, Saint Louis University","place":["St. Louis, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8535-7620","authenticated-orcid":true,"given":"Dapeng","family":"Zhang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01p7jjy08","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biology, College of Arts and Sciences, Saint Louis University","place":["St. Louis, USA"]},{"id":[{"id":"https:\/\/ror.org\/01p7jjy08","id-type":"ROR","asserted-by":"publisher"}],"name":"Program of Bioinformatics and Computational Biology, Saint Louis University","place":["St. Louis, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1503-170X","authenticated-orcid":true,"given":"Jue D.","family":"Wang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01y2jtd41","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Bacteriology, University of Wisconsin-Madison","place":["Madison, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4213-4050","authenticated-orcid":true,"given":"Mee-Ngan F.","family":"Yap","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02ets8c94","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology-Immunology, Northwestern University Feinberg School of Medicine","place":["Chicago, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41573-024-00897-5"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics11111570"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2411919121"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.3345\/cep.2019.00360"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.envint.2024.108680"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1101\/cshperspect.a025395"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.3109\/1040841X.2015.1136261"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.39.2.369"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12879-019-3779-8"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-25096-3"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41589-020-00715-0"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.26508\/lsa.202503325"},{"key":"e_1_3_4_14_2","unstructured":"Roberts MC. 2024. 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