{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:24:21Z","timestamp":1785403461153,"version":"3.56.0"},"posted":{"date-parts":[[2026,5,20]]},"group-title":"Microbiology","reference-count":77,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,5,20]],"date-time":"2026-05-20T00:00:00Z","timestamp":1779235200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"award":["R01 AI139074"],"award-info":[{"award-number":["R01 AI139074"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R21 AI142517"],"award-info":[{"award-number":["R21 AI142517"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,27]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Bacterial pathogens must adapt to dynamic host tissue environments to proliferate. Accordingly, elegant regulatory systems evolved to overcome challenges presented by the host and satisfy nutritional requirements. Sulfur is an essential macronutrient and Gram-positive bacteria such as\n                  <jats:italic>Staphylococcus aureus<\/jats:italic>\n                  balance this nutritional requirement by employing the transcriptional repressor, CymR. Previous investigations defined the\n                  <jats:italic>S. aureus<\/jats:italic>\n                  CymR regulon by comparing transcripts generated in a\n                  <jats:italic>cymR<\/jats:italic>\n                  mutant cultured in cystine replete, rich medium to wild type cells. This study defines the\n                  <jats:italic>S. aureus<\/jats:italic>\n                  CymR-dependent and -independent sulfur-starvation response in chemically defined growth conditions. Results demonstrate that the sulfur starvation and sulfur replete CymR regulons exhibit considerable overlap, including previously noted connections between iron acquisition, oxidative stress, and sulfur metabolism. The link between iron acquisition, oxidative stress, and sulfur metabolism is validated further by the finding that sulfur-containing glutathione (GSH) mitigates heme and peroxide toxicity. In addition to GSH, Cys and thiosulfate fulfill the\n                  <jats:italic>S. aureus<\/jats:italic>\n                  sulfur requirement. Transcriptional responses to organic (cysteine, cystine, reduced and oxidized GSH) or inorganic thiosulfate were quantified, revealing sulfur source-specific expression patterns. Thiosulfate induced the largest number of differentially expressed genes. Consequently, the thiosulfate transporter (SAUSA300_RS10985) has been confirmed as essential for\n                  <jats:italic>S. aureus<\/jats:italic>\n                  growth when thiosulfate is the sulfur source. Furthermore, we demonstrate that a hypothetical protein operonic with SAUSA300_RS10985, SAUSA300_RS10980, supports maximal growth on thiosulfate. Collectively, a resourceful transcriptomics framework is provided which underscores the dynamic nature of\n                  <jats:italic>S. aureus<\/jats:italic>\n                  sulfur metabolism.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>IMPORTANCE<\/jats:title>\n                  <jats:p>\n                    The opportunistic pathogen\n                    <jats:italic>Staphylococcus aureus<\/jats:italic>\n                    proliferates within diverse host environments and consequently is a leading cause of hospital-acquired infections. During colonization\n                    <jats:italic>S. aureus<\/jats:italic>\n                    must acquire essential nutrients like sulfur to survive. Regulation of machinery to procure host-derived metabolites occurs to satisfy nutritional requirements and maintain homeostasis to circumvent detrimental impacts on bacterial physiology. This report investigates the\n                    <jats:italic>S. aureus<\/jats:italic>\n                    regulatory response during various states of sulfur supplementation\n                    <jats:italic>in vitro<\/jats:italic>\n                    , enhancing our knowledge of staphylococcal sulfur metabolism and represents a repository of information that will guide future research surrounding\n                    <jats:italic>S. aureus<\/jats:italic>\n                    sulfur acquisition and metabolism.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.05.20.726469","type":"posted-content","created":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T05:25:17Z","timestamp":1779341117000},"source":"Crossref","is-referenced-by-count":0,"title":["Defining the transcriptional adaptation of\n                  <i>Staphylococcus aureus<\/i>\n                  to a range of nutritional sulfur supplementation"],"prefix":"10.64898","author":[{"given":"Paige J.","family":"Kies","sequence":"first","affiliation":[{"name":"Department of Microbiology, Genetics, & Immunology Michigan State University"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3485-2434","authenticated-orcid":false,"given":"Cristina","family":"Kraemer Zimpel","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Genetics, & Immunology Michigan State University"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joshua M.","family":"Lensmire","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Genetics, & Immunology Michigan State University"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"McKenna R.","family":"Major","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Genetics, & Immunology Michigan State University"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Troy A.","family":"Burtchett","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Genetics, & Immunology Michigan State University"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael R.","family":"Wischer","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Genetics, & Immunology Michigan State University"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6148-7850","authenticated-orcid":false,"given":"Neal D.","family":"Hammer","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Genetics, & Immunology Michigan State University"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026053012552943000_2026.05.20.726469v2.1","doi-asserted-by":"publisher","DOI":"10.1001\/jama.298.15.1763"},{"key":"2026053012552943000_2026.05.20.726469v2.2","doi-asserted-by":"crossref","unstructured":"Lakhundi S , Zhang K . 2018. Methicillin-Resistant Staphylococcus aureus: Molecular Characterization, Evolution, and Epidemiology. Clin Microbiol Rev 31.","DOI":"10.1128\/CMR.00020-18"},{"key":"2026053012552943000_2026.05.20.726469v2.3","doi-asserted-by":"crossref","first-page":"821","DOI":"10.1016\/j.tim.2020.04.004","article-title":"Genetic Regulation of Metal Ion Homeostasis in Staphylococcus aureus","volume":"28","year":"2020","journal-title":"Trends Microbiol"},{"key":"2026053012552943000_2026.05.20.726469v2.4","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.micro.57.030502.090938"},{"key":"2026053012552943000_2026.05.20.726469v2.5","doi-asserted-by":"publisher","DOI":"10.1039\/c2mt20142a"},{"key":"2026053012552943000_2026.05.20.726469v2.6","doi-asserted-by":"publisher","DOI":"10.1016\/j.freeradbiomed.2014.11.013"},{"key":"2026053012552943000_2026.05.20.726469v2.7","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.2002.02851.x"},{"key":"2026053012552943000_2026.05.20.726469v2.8","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1002264"},{"key":"2026053012552943000_2026.05.20.726469v2.9","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2015.05.014"},{"key":"2026053012552943000_2026.05.20.726469v2.10","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.biochem.74.082803.133518"},{"key":"2026053012552943000_2026.05.20.726469v2.11","doi-asserted-by":"publisher","DOI":"10.1111\/mmi.12460"},{"key":"2026053012552943000_2026.05.20.726469v2.12","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbpa.2008.01.021"},{"key":"2026053012552943000_2026.05.20.726469v2.13","doi-asserted-by":"crossref","unstructured":"Nagy P , Becker JD , Mallo RC , Ashby MT . 2007. The Jekyll and Hyde Roles of Cysteine Derivatives during Oxidative Stress, p 193-212, New Biocides Development, vol 967. American Chemical Society.","DOI":"10.1021\/bk-2007-0967.ch010"},{"key":"2026053012552943000_2026.05.20.726469v2.14","doi-asserted-by":"publisher","DOI":"10.1128\/JB.185.6.1942-1950.2003"},{"key":"2026053012552943000_2026.05.20.726469v2.15","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.RA120.014814"},{"key":"2026053012552943000_2026.05.20.726469v2.16","doi-asserted-by":"crossref","first-page":"e0057921","DOI":"10.1128\/iai.00579-21","article-title":"A Resourceful Race: Bacterial Scavenging of Host Sulfur Metabolism during Colonization","volume":"90","year":"2022","journal-title":"Infect Immun"},{"key":"2026053012552943000_2026.05.20.726469v2.17","doi-asserted-by":"publisher","DOI":"10.1128\/JB.188.6.2184-2197.2006"},{"key":"2026053012552943000_2026.05.20.726469v2.18","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2009.06760.x"},{"key":"2026053012552943000_2026.05.20.726469v2.19","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.359737"},{"key":"2026053012552943000_2026.05.20.726469v2.20","doi-asserted-by":"crossref","first-page":"e0037425","DOI":"10.1128\/mbio.00374-25","article-title":"Analysis of genetic requirements and nutrient availability for Staphylococcus aureus growth in cystic fibrosis sputum","volume":"16","year":"2025","journal-title":"mBio"},{"key":"2026053012552943000_2026.05.20.726469v2.21","doi-asserted-by":"publisher","DOI":"10.1128\/JB.186.6.1579-1590.2004"},{"key":"2026053012552943000_2026.05.20.726469v2.22","doi-asserted-by":"crossref","unstructured":"Lensmire JM , Dodson JP , Hsueh BY , Wischer MR , Delekta PC , Shook JC , Ottosen EN , Kies PJ , Ravi J , Hammer ND . 2020. The Staphylococcus aureus Cystine Transporters TcyABC and TcyP Facilitate Nutrient Sulfur Acquisition during Infection. Infect Immun 88.","DOI":"10.1128\/IAI.00690-19"},{"key":"2026053012552943000_2026.05.20.726469v2.23","doi-asserted-by":"crossref","first-page":"e1010834","DOI":"10.1371\/journal.pgen.1010834","article-title":"The glutathione import system satisfies the Staphylococcus aureus nutrient sulfur requirement and promotes interspecies competition","volume":"19","year":"2023","journal-title":"PLoS Genet"},{"key":"2026053012552943000_2026.05.20.726469v2.24","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1000894"},{"key":"2026053012552943000_2026.05.20.726469v2.25","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.aba7637"},{"key":"2026053012552943000_2026.05.20.726469v2.26","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.3002601"},{"key":"2026053012552943000_2026.05.20.726469v2.27","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1128\/jb.124.1.201-211.1975","article-title":"Transformation analysis of three linkage groups in Staphylococcus aureus","volume":"124","year":"1975","journal-title":"J Bacteriol"},{"key":"2026053012552943000_2026.05.20.726469v2.28","doi-asserted-by":"crossref","unstructured":"Vitko NP , Richardson AR . 2013. Laboratory maintenance of methicillin-resistant Staphylococcus aureus (MRSA). Curr Protoc Microbiol Chapter 9:Unit 9C 2.","DOI":"10.1002\/9780471729259.mc09c02s28"},{"key":"2026053012552943000_2026.05.20.726469v2.29","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jafc.7b02391"},{"key":"2026053012552943000_2026.05.20.726469v2.30","doi-asserted-by":"publisher","DOI":"10.1089\/ars.2005.7.813"},{"key":"2026053012552943000_2026.05.20.726469v2.31","doi-asserted-by":"crossref","first-page":"656","DOI":"10.3390\/cryst9120656","article-title":"Crystal Structure of Bacterial Cystathionine \u0393-Lyase in The Cysteine Biosynthesis Pathway of Staphylococcus aureus","volume":"9","year":"2019","journal-title":"Crystals"},{"key":"2026053012552943000_2026.05.20.726469v2.32","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01273-06"},{"key":"2026053012552943000_2026.05.20.726469v2.33","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M805951200"},{"key":"2026053012552943000_2026.05.20.726469v2.34","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.69.6.3744-3754.2001"},{"key":"2026053012552943000_2026.05.20.726469v2.35","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.0020087"},{"key":"2026053012552943000_2026.05.20.726469v2.36","doi-asserted-by":"publisher","DOI":"10.1128\/JB.183.2.468-475.2001"},{"key":"2026053012552943000_2026.05.20.726469v2.37","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2008413117"},{"key":"2026053012552943000_2026.05.20.726469v2.38","doi-asserted-by":"publisher","DOI":"10.1023\/A:1026650023400"},{"key":"2026053012552943000_2026.05.20.726469v2.39","doi-asserted-by":"publisher","DOI":"10.1089\/ars.2017.7354"},{"key":"2026053012552943000_2026.05.20.726469v2.40","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijmm.2017.11.011"},{"key":"2026053012552943000_2026.05.20.726469v2.41","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.69.2.885-896.2001"},{"key":"2026053012552943000_2026.05.20.726469v2.42","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2007.05608.x"},{"key":"2026053012552943000_2026.05.20.726469v2.43","doi-asserted-by":"crossref","first-page":"1351","DOI":"10.1016\/j.chembiol.2016.09.012","article-title":"A Small-Molecule Inhibitor of Iron-Sulfur Cluster Assembly Uncovers a Link between Virulence Regulation and Metabolism in Staphylococcus aureus","volume":"23","year":"2016","journal-title":"Cell Chem Biol"},{"key":"2026053012552943000_2026.05.20.726469v2.44","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbpa.2011.02.012"},{"key":"2026053012552943000_2026.05.20.726469v2.45","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01393-12"},{"key":"2026053012552943000_2026.05.20.726469v2.46","doi-asserted-by":"crossref","unstructured":"Stewart LJ , Ong CY , Zhang MM , Brouwer S , McIntyre L , Davies MR , Walker MJ , McEwan AG , Waldron KJ , Djoko KY . 2020. Role of Glutathione in Buffering Excess Intracellular Copper in Streptococcus pyogenes . mBio 11.","DOI":"10.1128\/mBio.02804-20"},{"key":"2026053012552943000_2026.05.20.726469v2.47","doi-asserted-by":"publisher","DOI":"10.1016\/j.bcmd.2015.09.003"},{"key":"2026053012552943000_2026.05.20.726469v2.48","doi-asserted-by":"publisher","DOI":"10.1126\/science.1081147"},{"key":"2026053012552943000_2026.05.20.726469v2.49","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.bi.52.070183.003431"},{"key":"2026053012552943000_2026.05.20.726469v2.50","doi-asserted-by":"publisher","DOI":"10.1089\/ars.2006.8.753"},{"key":"2026053012552943000_2026.05.20.726469v2.51","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2007.03.001"},{"key":"2026053012552943000_2026.05.20.726469v2.52","doi-asserted-by":"publisher","DOI":"10.1128\/jb.00337-23"},{"key":"2026053012552943000_2026.05.20.726469v2.53","doi-asserted-by":"crossref","unstructured":"Joo HS , Chatterjee SS , Villaruz AE , Dickey SW , Tan VY , Chen Y , Sturdevant DE , Ricklefs SM , Otto M . 2016. Mechanism of Gene Regulation by a Staphylococcus aureus Toxin. mBio 7.","DOI":"10.1128\/mBio.01579-16"},{"key":"2026053012552943000_2026.05.20.726469v2.54","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.049692-0"},{"key":"2026053012552943000_2026.05.20.726469v2.55","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00491-10"},{"key":"2026053012552943000_2026.05.20.726469v2.56","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.50.1.336-343.2006"},{"key":"2026053012552943000_2026.05.20.726469v2.57","doi-asserted-by":"publisher","DOI":"10.1099\/00221287-147-5-1259"},{"key":"2026053012552943000_2026.05.20.726469v2.58","doi-asserted-by":"publisher","DOI":"10.1111\/mmi.12835"},{"key":"2026053012552943000_2026.05.20.726469v2.59","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.02370-16"},{"key":"2026053012552943000_2026.05.20.726469v2.60","first-page":"307","article-title":"Defect of RNA pyrophosphohydrolase RppH enhances fermentative production of L-cysteine in Escherichia coli","volume":"66","year":"2021","journal-title":"J Gen Appl Microbiol"},{"key":"2026053012552943000_2026.05.20.726469v2.61","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462-2920.2010.02210.x"},{"key":"2026053012552943000_2026.05.20.726469v2.62","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01921-07"},{"key":"2026053012552943000_2026.05.20.726469v2.63","doi-asserted-by":"publisher","DOI":"10.1016\/0006-2952(87)90441-2"},{"key":"2026053012552943000_2026.05.20.726469v2.64","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.270.42.24876"},{"key":"2026053012552943000_2026.05.20.726469v2.65","doi-asserted-by":"publisher","DOI":"10.1016\/j.freeradbiomed.2013.08.002"},{"key":"2026053012552943000_2026.05.20.726469v2.66","doi-asserted-by":"publisher","DOI":"10.1128\/jb.172.6.3351-3357.1990"},{"key":"2026053012552943000_2026.05.20.726469v2.67","doi-asserted-by":"publisher","DOI":"10.1128\/jb.177.14.4134-4136.1995"},{"key":"2026053012552943000_2026.05.20.726469v2.68","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00537-12"},{"key":"2026053012552943000_2026.05.20.726469v2.69","doi-asserted-by":"publisher","DOI":"10.1016\/0042-6822(67)90105-5"},{"key":"2026053012552943000_2026.05.20.726469v2.70","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-59745-321-9_7"},{"key":"2026053012552943000_2026.05.20.726469v2.71","doi-asserted-by":"publisher","DOI":"10.1128\/JB.186.6.1838-1850.2004"},{"key":"2026053012552943000_2026.05.20.726469v2.72","doi-asserted-by":"publisher","DOI":"10.1016\/j.mimet.2009.02.011"},{"key":"2026053012552943000_2026.05.20.726469v2.73","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-014-0550-8"},{"key":"2026053012552943000_2026.05.20.726469v2.74","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gky1085"},{"key":"2026053012552943000_2026.05.20.726469v2.75","doi-asserted-by":"publisher","DOI":"10.1101\/2021.06.03.446934"},{"key":"2026053012552943000_2026.05.20.726469v2.76","doi-asserted-by":"publisher","DOI":"10.1093\/bib\/bbx085"},{"key":"2026053012552943000_2026.05.20.726469v2.77","doi-asserted-by":"crossref","unstructured":"Austin CM , Garabaglu S , Krute CN , Ridder MJ , Seawell NA , Markiewicz MA , Boyd JM , Bose JL . 2019. Contribution of YjbIH to Virulence Factor Expression and Host Colonization in Staphylococcus aureus . Infect Immun 87.","DOI":"10.1128\/IAI.00155-19"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.05.20.726469","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,30]],"date-time":"2026-05-30T19:55:39Z","timestamp":1780170939000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.05.20.726469"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,20]]},"references-count":77,"URL":"https:\/\/doi.org\/10.64898\/2026.05.20.726469","relation":{},"subject":[],"published":{"date-parts":[[2026,5,20]]},"subtype":"preprint"}}