{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T20:00:11Z","timestamp":1768507211514,"version":"3.49.0"},"posted":{"date-parts":[[2023,7,24]]},"group-title":"Microbiology","reference-count":44,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,7,24]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  <jats:italic>Staphylococcus aureus<\/jats:italic>\n                  and\n                  <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n                  are the most common bacterial pathogens isolated from cystic fibrosis (CF) related lung infections. When both of these opportunistic pathogens are found in a coinfection, CF patients tend to have higher rates of pulmonary exacerbations and experience a more rapid decrease in lung function. When cultured together under standard laboratory conditions, it is often observed that\n                  <jats:italic>P. aeruginosa<\/jats:italic>\n                  effectively inhibits\n                  <jats:italic>S. aureus<\/jats:italic>\n                  growth. Previous work from our group revealed that\n                  <jats:italic>S. aureus<\/jats:italic>\n                  from CF infections have isolate-specific survival capabilities when cocultured with\n                  <jats:italic>P. aeruginosa<\/jats:italic>\n                  . In this study, we designed a serial transfer evolution experiment to identify mutations that allow\n                  <jats:italic>S. aureus<\/jats:italic>\n                  to adapt to the presence of\n                  <jats:italic>P. aeruginosa<\/jats:italic>\n                  . Using\n                  <jats:italic>S. aureus<\/jats:italic>\n                  USA300 JE2 as our ancestral strain, populations of\n                  <jats:italic>S. aureus<\/jats:italic>\n                  were repeatedly cocultured with fresh\n                  <jats:italic>P. aeruginosa<\/jats:italic>\n                  strain, PAO1. After 8 coculture periods,\n                  <jats:italic>S. aureus<\/jats:italic>\n                  populations that survived better in the presence of PAO1 were observed. We found two independent mutations in the highly conserved\n                  <jats:italic>S. aureus<\/jats:italic>\n                  aspartate transporter,\n                  <jats:italic>gltT<\/jats:italic>\n                  , that were unique to evolved\n                  <jats:italic>P. aeruginosa<\/jats:italic>\n                  -tolerant isolates. Subsequent phenotypic testing demonstrated that\n                  <jats:italic>gltT<\/jats:italic>\n                  mutants have reduced uptake of glutamate and outcompete wild-type\n                  <jats:italic>S. aureus<\/jats:italic>\n                  when glutamate is absent from chemically-defined media. These findings together demonstrate that the presence of\n                  <jats:italic>P. aeruginosa<\/jats:italic>\n                  exerts selective pressure on\n                  <jats:italic>S. aureus<\/jats:italic>\n                  to alter its uptake and metabolism of key amino acids when the two bacteria are cultured together.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Importance<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Staphylococcus aureus<\/jats:italic>\n                    and\n                    <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n                    are the two most common bacterial pathogens that infect people with the genetic disease, cystic fibrosis (CF). They are often found together in CF-associated polymicrobial infections that are associated with worse patient prognosis. Understanding how these very different opportunistic pathogens influence each other in a shared environment is pertinent to improving the treatment of polymicrobial infections. While much attention has been brought to the interspecific interactions between\n                    <jats:italic>S. aureus<\/jats:italic>\n                    and\n                    <jats:italic>P. aeruginosa,<\/jats:italic>\n                    few studies have used experimental evolution methods to identify determinants of their competition and coexistence. Here, we use a serial transfer experimental evolution approach and identified a single genetic change associated with improved survival of\n                    <jats:italic>S. aureus<\/jats:italic>\n                    in the presence of\n                    <jats:italic>P. aeruginosa.<\/jats:italic>\n                    Our findings implicate metabolism of shared resources as an important factor in\n                    <jats:italic>S. aureus\u2019s<\/jats:italic>\n                    ability to survive in the presence of\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    .\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2023.07.24.550428","type":"posted-content","created":{"date-parts":[[2023,7,25]],"date-time":"2023-07-25T00:55:15Z","timestamp":1690246515000},"source":"Crossref","is-referenced-by-count":0,"title":["Experimentally Evolved\n                  <i>Staphylococcus aureus<\/i>\n                  Survives in the Presence of\n                  <i>Pseudomonas aeruginosa<\/i>\n                  by Acquiring Mutations in the Amino Acid Transporter, GltT"],"prefix":"10.64898","author":[{"given":"Ashley M.","family":"Alexander","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Justin M.","family":"Luu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vishnu","family":"Raghuram","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giulia","family":"Bottacin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Simon","family":"van Vliet","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8966-9680","authenticated-orcid":false,"given":"Timothy D.","family":"Read","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5285-5188","authenticated-orcid":false,"given":"Joanna B.","family":"Goldberg","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2023073023402188000_2023.07.24.550428v1.1","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00047-21"},{"issue":"12","key":"2023073023402188000_2023.07.24.550428v1.2","doi-asserted-by":"crossref","first-page":"1226","DOI":"10.1038\/s41592-019-0582-9","article-title":"ilastik: interactive machine learning for (bio)image analysis","volume":"16","year":"2019","journal-title":"Nature Methods"},{"key":"2023073023402188000_2023.07.24.550428v1.3","first-page":"3","article-title":"Genotypic and Phenotypic Diversity of Staphylococcus aureus Isolates from Cystic Fibrosis Patient Lung Infections and Their Interactions with Pseudomonas aeruginosa","volume":"11","year":"2020","journal-title":"MBio"},{"issue":"4","key":"2023073023402188000_2023.07.24.550428v1.4","first-page":"e00976","article-title":"Staphylococcus aureus and Pseudomonas aeruginosa Isolates from the Same Cystic Fibrosis Respiratory Sample Coexist in Coculture","volume":"10","year":"2022","journal-title":"Microbiology Spectrum"},{"key":"2023073023402188000_2023.07.24.550428v1.5","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btu170"},{"key":"2023073023402188000_2023.07.24.550428v1.6","doi-asserted-by":"publisher","DOI":"10.1016\/j.tim.2012.04.005"},{"key":"2023073023402188000_2023.07.24.550428v1.7","doi-asserted-by":"publisher","DOI":"10.1111\/j.1461-0248.2007.01125.x"},{"key":"2023073023402188000_2023.07.24.550428v1.8","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0603072103"},{"issue":"12","key":"2023073023402188000_2023.07.24.550428v1.9","doi-asserted-by":"crossref","first-page":"3093","DOI":"10.1038\/s41396-020-00741-9","article-title":"Trophic cooperation promotes bacterial survival of Staphylococcus aureus and Pseudomonas aeruginosa","volume":"14","year":"2020","journal-title":"The ISME Journal"},{"key":"2023073023402188000_2023.07.24.550428v1.10","doi-asserted-by":"publisher","DOI":"10.1016\/j.plasmid.2014.08.001"},{"key":"2023073023402188000_2023.07.24.550428v1.11","unstructured":"Cystic Fibrosis Foundation. 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