{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T16:07:29Z","timestamp":1753891649952,"version":"3.41.2"},"reference-count":55,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,6,3]],"date-time":"2022-06-03T00:00:00Z","timestamp":1654214400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100008519","name":"University of Arkansas for Medical Sciences","doi-asserted-by":"publisher","award":["Z1-50328-01"],"award-info":[{"award-number":["Z1-50328-01"]}],"id":[{"id":"10.13039\/100008519","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:p><jats:italic>Staphylococcus aureus<\/jats:italic> is an important human pathogen that can infect almost every organ system, resulting in a high incidence of morbidity and mortality. The <jats:italic>msaABCR<\/jats:italic> operon is an important regulator of several staphylococcal phenotypes, including biofilm development, cell wall crosslinking, antibiotic resistance, oxidative stress, and acute and chronic implant-associated osteomyelitis. Our previous study showed that, by modulating murein hydrolase activity, the <jats:italic>msaABCR<\/jats:italic> operon negatively regulates the proteases that govern cell death. Here, we report further elucidation of the mechanism of cell death, which is regulated by the <jats:italic>msaABCR<\/jats:italic> operon at the molecular level in the USA300 LAC strain. We showed that deletion of <jats:italic>msaABCR<\/jats:italic> enhances weak-acid-dependent cell death, because, in the biofilm microenvironment, this mutant strain consumes glucose and produces acetate and acetoin at higher rates than wild-type USA300 LAC strain. We proposed the increased intracellular acidification leads to increased cell death. MsaB, a dual-function transcription factor and RNA chaperone, is a negative regulator of the <jats:italic>cidR<\/jats:italic> regulon, which has been shown to play an important role in overflow metabolism and programmed cell death during biofilm development in <jats:italic>S. aureus<\/jats:italic>. We found that MsaB binds directly to the <jats:italic>cidR<\/jats:italic> promoter, which represses expression of the <jats:italic>cidR<\/jats:italic> regulon and prevents transcription of the <jats:italic>cidABC<\/jats:italic> and <jats:italic>alsSD<\/jats:italic> operons. In addition, we observed that pyruvate induced expression of the <jats:italic>msaABCR<\/jats:italic> operon (MsaB). The results reported here have enabled us to decipher the role of the <jats:italic>msaABCR<\/jats:italic> operon in staphylococcal metabolic adaption during biofilm development.<\/jats:p>","DOI":"10.3389\/fmicb.2022.914512","type":"journal-article","created":{"date-parts":[[2022,6,3]],"date-time":"2022-06-03T09:30:10Z","timestamp":1654248610000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":6,"title":["Delineating the Role of the msaABCR Operon in Staphylococcal Overflow Metabolism"],"prefix":"10.3389","volume":"13","author":[{"given":"Bibek","family":"G C","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gyan S.","family":"Sahukhal","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mohamed O.","family":"Elasri","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2022,6,3]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"309","DOI":"10.1111\/mmi.14418","article-title":"Analysis of the CodY RNome reveals RsaD as a stress-responsive riboregulator of overflow metabolism in Staphylococcus aureus","volume":"113","author":"Augagneur","year":"2020","journal-title":"Mol. Microbiol."},{"key":"ref2","doi-asserted-by":"publisher","first-page":"58","DOI":"10.1016\/j.plasmid.2005.05.005","article-title":"Allelic replacement in Staphylococcus aureus with inducible counter-selection","volume":"55","author":"Bae","year":"2006","journal-title":"Plasmid"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"e00294-18","DOI":"10.1128\/JB.00294-18","article-title":"MsaB and CodY interact to regulate Staphylococcus aureus capsule in a nutrient-dependent manner","volume":"200","author":"Batte","year":"2018","journal-title":"J. Bacteriol."},{"key":"ref4","doi-asserted-by":"publisher","first-page":"575","DOI":"10.1099\/mic.0.000243","article-title":"MsaB activates capsule production at the transcription level in Staphylococcus aureus","volume":"162","author":"Batte","year":"2016","journal-title":"Microbiology"},{"key":"ref5","doi-asserted-by":"publisher","first-page":"721","DOI":"10.1038\/nrmicro1743","article-title":"The biological role of death and lysis in biofilm development","volume":"5","author":"Bayles","year":"2007","journal-title":"Nat. Rev. Microbiol."},{"key":"ref6","doi-asserted-by":"publisher","first-page":"942","DOI":"10.1111\/mmi.13433","article-title":"The LysR-type transcriptional regulator, CidR, regulates stationary phase cell death in Staphylococcus aureus","volume":"101","author":"Chaudhari","year":"2016","journal-title":"Mol. Microbiol."},{"key":"ref7","doi-asserted-by":"publisher","first-page":"29440","DOI":"10.1074\/jbc.M113.502039","article-title":"Secreted proteases control autolysin-mediated biofilm growth of Staphylococcus aureus","volume":"288","author":"Chen","year":"2013","journal-title":"J. Biol. Chem."},{"key":"ref8","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1111\/j.1524-475X.2007.00303.x","article-title":"Microscopic and physiologic evidence for biofilm-associated wound colonization in vivo","volume":"16","author":"Davis","year":"2008","journal-title":"Wound Repair Regen."},{"key":"ref9","doi-asserted-by":"publisher","first-page":"1373","DOI":"10.1080\/14760584.2016.1179583","article-title":"Staphylococcus aureus: the current state of disease, pathophysiology and strategies for prevention","volume":"15","author":"Dayan","year":"2016","journal-title":"Expert Rev. Vaccines"},{"key":"ref10","article-title":"Identification and Characterization of msaB gene Involved in Biofilm Formation and Virulence in Staphylococcus aureus","volume":"49","author":"Elbarasi","year":"2014","journal-title":"master\u2019s theses."},{"key":"ref11","doi-asserted-by":"publisher","first-page":"393","DOI":"10.1016\/j.mimet.2012.01.007","article-title":"Comparison of in vitro and in vivo systems to study Ica-independent Staphylococcus aureus biofilms","volume":"88","author":"Ferreira","year":"2012","journal-title":"J. Microbiol. Methods"},{"key":"ref12","doi-asserted-by":"publisher","first-page":"e00680-00619","DOI":"10.1128\/AAC.00680-19","article-title":"Role of the msaABCR operon in cell wall biosynthesis, autolysis, integrity, and antibiotic resistance in Staphylococcus aureus","volume":"63","author":"GC","year":"2019","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref13","doi-asserted-by":"publisher","first-page":"1304","DOI":"10.1128\/IAI.68.3.1304-1311.2000","article-title":"Direct quantitative transcript analysis of the agr regulon of Staphylococcus aureus during human infection in comparison to the expression profile in vitro","volume":"68","author":"Goerke","year":"2000","journal-title":"Infect. Immun."},{"key":"ref15","doi-asserted-by":"publisher","first-page":"e02272-17","DOI":"10.1128\/mBio.02272-17","article-title":"Staphylococcus aureus responds to the central metabolite pyruvate to regulate virulence","volume":"9","author":"Harper","year":"2018","journal-title":"MBio"},{"key":"ref16","doi-asserted-by":"publisher","first-page":"225","DOI":"10.1016\/j.micpath.2017.07.004","article-title":"Vancomycin-induced biofilm formation by methicillin-resistant Staphylococcus aureus is associated with the secretion of membrane vesicles","volume":"110","author":"He","year":"2017","journal-title":"Microb. Pathog."},{"key":"ref17","doi-asserted-by":"publisher","first-page":"236","DOI":"10.1111\/j.1574-695X.2011.00846.x","article-title":"Vancomycin promotes the bacterial autolysis, release of extracellular DNA, and biofilm formation in vancomycin-non-susceptible Staphylococcus aureus","volume":"63","author":"Hsu","year":"2011","journal-title":"FEMS Immunol. Med. Microbiol."},{"key":"ref18","doi-asserted-by":"publisher","first-page":"e0134852","DOI":"10.1371\/journal.pone.0134852","article-title":"High glucose concentration promotes Vancomycin-enhanced biofilm formation of Vancomycin-non-susceptible Staphylococcus aureus in diabetic mice","volume":"10","author":"Hsu","year":"2015","journal-title":"PLoS One"},{"key":"ref20","doi-asserted-by":"publisher","first-page":"e00696","DOI":"10.1128\/mBio.00696-13","article-title":"The Staphylococcus aureus SrrAB two-component system promotes resistance to nitrosative stress and hypoxia","volume":"4","author":"Kinkel","year":"2013","journal-title":"MBio"},{"key":"ref21","doi-asserted-by":"publisher","first-page":"1205","DOI":"10.1021\/acs.jproteome.5b01089","article-title":"Redox imbalance underlies the fitness defect associated with inactivation of the Pta-AckA pathway in Staphylococcus aureus","volume":"15","author":"Marshall","year":"2016","journal-title":"J. Proteome Res."},{"key":"ref23","doi-asserted-by":"publisher","first-page":"191","DOI":"10.1002\/jobm.201000221","article-title":"Effect of sub-lethal doses of vancomycin and oxacillin on biofilm formation by vancomycin intermediate resistant Staphylococcus aureus","volume":"51","author":"Mirani","year":"2011","journal-title":"J. Basic Microbiol."},{"key":"ref24","doi-asserted-by":"publisher","first-page":"365","DOI":"10.1111\/mmi.13634","article-title":"Staphylococcus aureus biofilm: a complex developmental organism","volume":"104","author":"Moormeier","year":"2017","journal-title":"Mol. Microbiol."},{"key":"ref25","doi-asserted-by":"publisher","first-page":"6832","DOI":"10.1128\/AEM.00881-08","article-title":"The Bacillus subtilis ydjL (bdhA) gene encodes acetoin reductase\/2,3-butanediol dehydrogenase","volume":"74","author":"Nicholson","year":"2008","journal-title":"Appl. Environ. Microbiol."},{"key":"ref26","doi-asserted-by":"publisher","first-page":"6","DOI":"10.1128\/microbiolspec.GPP3-0023-2018","article-title":"Staphylococcal biofilms","volume":"6","author":"Otto","year":"2018","journal-title":"Microbiol. Spectr."},{"key":"ref27","doi-asserted-by":"publisher","first-page":"e00417-19","DOI":"10.1128\/JB.00417-19","article-title":"The msaABCR operon regulates the response to oxidative stress in Staphylococcus aureus","volume":"201","author":"Pandey","year":"2019","journal-title":"J. Bacteriol."},{"key":"ref001","doi-asserted-by":"publisher","first-page":"657753","DOI":"10.3389\/fmicb.2021.657753","article-title":"The msaABCR Operon Regulates Persister Formation by Modulating Energy Metabolism in Staphylococcus aureus","volume":"12","author":"Pandey","year":"2021","journal-title":"Front. Microbiol."},{"key":"ref28","doi-asserted-by":"publisher","first-page":"1664","DOI":"10.1111\/j.1365-2958.2005.04653.x","article-title":"The Staphylococcus aureus cidC gene encodes a pyruvate oxidase that affects acetate metabolism and cell death in stationary phase","volume":"56","author":"Patton","year":"2005","journal-title":"Mol. Microbiol."},{"key":"ref29","doi-asserted-by":"publisher","first-page":"1395","DOI":"10.1111\/j.1365-2958.2006.05034.x","article-title":"The role of proton motive force in expression of the Staphylococcus aureus cid and lrg operons","volume":"59","author":"Patton","year":"2006","journal-title":"Mol. Microbiol."},{"key":"ref30","doi-asserted-by":"publisher","first-page":"413","DOI":"10.1016\/S0891-5520(01)00003-4","article-title":"Staphylococcus aureus bacteremia and endocarditis","volume":"16","author":"Petti","year":"2002","journal-title":"Infect. Dis. Clin. North Am."},{"key":"ref31","doi-asserted-by":"publisher","first-page":"729","DOI":"10.1046\/j.1365-2958.2003.t01-1-03720.x","article-title":"Death's toolbox: examining the molecular components of bacterial programmed cell death","volume":"50","author":"Rice","year":"2003","journal-title":"Mol. Microbiol."},{"key":"ref32","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1128\/MMBR.00030-07","article-title":"Molecular control of bacterial death and lysis","volume":"72","author":"Rice","year":"2008","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref33","doi-asserted-by":"publisher","first-page":"2635","DOI":"10.1128\/JB.185.8.2635-2643.2003","article-title":"The Staphylococcus aureus cidAB operon: evaluation of its role in regulation of murein hydrolase activity and penicillin tolerance","volume":"185","author":"Rice","year":"2003","journal-title":"J. Bacteriol."},{"key":"ref34","doi-asserted-by":"publisher","first-page":"813","DOI":"10.1128\/JB.187.3.813-821.2005","article-title":"Acetic acid induces expression of the Staphylococcus aureus cidABC and lrgAB murein hydrolase regulator operons","volume":"187","author":"Rice","year":"2005","journal-title":"J. Bacteriol."},{"key":"ref35","doi-asserted-by":"publisher","first-page":"3029","DOI":"10.1128\/JB.186.10.3029-3037.2004","article-title":"Transcription of the Staphylococcus aureus cid and lrg murein hydrolase regulators is affected by sigma factor B","volume":"186","author":"Rice","year":"2004","journal-title":"J. Bacteriol."},{"key":"ref36","doi-asserted-by":"publisher","first-page":"159","DOI":"10.1128\/IAI.69.1.159-169.2001","article-title":"Description of staphylococcus serine protease (ssp) operon in Staphylococcus aureus and nonpolar inactivation of sspA-encoded serine protease","volume":"69","author":"Rice","year":"2001","journal-title":"Infect. Immun."},{"key":"ref38","doi-asserted-by":"publisher","first-page":"e00530-19","DOI":"10.1128\/IAI.00530-19","article-title":"The impacts of msaABCR on sarA-associated phenotypes are different in divergent clinical isolates of Staphylococcus aureus","volume":"8","author":"Rom","year":"2020","journal-title":"Infect. Immun."},{"key":"ref39","doi-asserted-by":"publisher","first-page":"211","DOI":"10.1016\/j.mib.2011.12.010","article-title":"The control of death and lysis in staphylococcal biofilms: a coordination of physiological signals","volume":"15","author":"Sadykov","year":"2012","journal-title":"Curr. Opin. Microbiol."},{"key":"ref40","doi-asserted-by":"publisher","first-page":"3458","DOI":"10.1099\/mic.0.051243-0","article-title":"CcpA coordinates central metabolism and biofilm formation in Staphylococcus epidermidis","volume":"157","author":"Sadykov","year":"2011","journal-title":"Microbiology"},{"key":"ref41","doi-asserted-by":"publisher","first-page":"3035","DOI":"10.1128\/JB.00042-13","article-title":"Inactivation of the Pta-AckA pathway causes cell death in Staphylococcus aureus","volume":"195","author":"Sadykov","year":"2013","journal-title":"J. Bacteriol."},{"key":"ref42","doi-asserted-by":"publisher","first-page":"e00371","DOI":"10.1128\/JB.00371-19","article-title":"CidR and CcpA synergistically regulate Staphylococcus aureus cidABC expression","volume":"201","author":"Sadykov","year":"2019","journal-title":"J. Bacteriol."},{"key":"ref43","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1093\/femsle\/fnv006","article-title":"msaABCR operon positively regulates biofilm development by repressing proteases and autolysis in Staphylococcus aureus","volume":"362","author":"Sahukhal","year":"2015","journal-title":"FEMS Microbiol. Lett."},{"key":"ref44","doi-asserted-by":"publisher","first-page":"154","DOI":"10.1186\/1471-2180-14-154","article-title":"Identification and characterization of an operon, msaABCR, that controls virulence and biofilm development in Staphylococcus aureus","volume":"14","author":"Sahukhal","year":"2014","journal-title":"BMC Microbiol."},{"key":"ref45","doi-asserted-by":"publisher","first-page":"218","DOI":"10.1186\/s12866-017-1129-9","article-title":"msaABCR operon is involved in persister cell formation in Staphylococcus aureus","volume":"17","author":"Sahukhal","year":"2017","journal-title":"BMC Microbiol."},{"key":"ref46","doi-asserted-by":"publisher","first-page":"324","DOI":"10.1186\/s12866-020-01964-8","article-title":"The role of the msaABCR operon in implant-associated chronic osteomyelitis in Staphylococcus aureus USA300 LAC","volume":"20","author":"Sahukhal","year":"2020","journal-title":"BMC Microbiol."},{"key":"ref47","doi-asserted-by":"publisher","first-page":"6685","DOI":"10.1128\/AAC.03280-14","article-title":"The msaABCR operon regulates resistance in vancomycin-intermediate Staphylococcus aureus strains","volume":"58","author":"Samanta","year":"2014","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref48","doi-asserted-by":"publisher","first-page":"221","DOI":"10.1186\/1471-2180-8-221","article-title":"The role of msa in Staphylococcus aureus biofilm formation","volume":"8","author":"Sambanthamoorthy","year":"2008","journal-title":"BMC Microbiol."},{"key":"ref49","doi-asserted-by":"publisher","first-page":"95","DOI":"10.1186\/1471-2180-9-95","article-title":"Effect of a glucose impulse on the CcpA regulon in Staphylococcus aureus","volume":"9","author":"Seidl","year":"2009","journal-title":"BMC Microbiol."},{"key":"ref50","doi-asserted-by":"publisher","first-page":"6373","DOI":"10.1128\/IAI.70.11.6373-6382.2002","article-title":"Staphylococcus aureus aconitase inactivation unexpectedly inhibits post-exponential-phase growth and enhances stationary-phase survival","volume":"70","author":"Somerville","year":"2002","journal-title":"Infect. Immun."},{"key":"ref51","doi-asserted-by":"publisher","first-page":"4724","DOI":"10.1128\/IAI.71.8.4724-4732.2003","article-title":"Correlation of acetate catabolism and growth yield in Staphylococcus aureus: implications for host-pathogen interactions","volume":"71","author":"Somerville","year":"2003","journal-title":"Infect. Immun."},{"key":"ref52","doi-asserted-by":"publisher","first-page":"e1004205","DOI":"10.1371\/journal.ppat.1004205","article-title":"A central role for carbon-overflow pathways in the modulation of bacterial cell death","volume":"10","author":"Thomas","year":"2014","journal-title":"PLoS Pathog."},{"key":"ref53","doi-asserted-by":"publisher","first-page":"5690","DOI":"10.1128\/JB.00314-08","article-title":"Regulation of autolysis-dependent extracellular DNA release by Enterococcus faecalis extracellular proteases influences biofilm development","volume":"190","author":"Thomas","year":"2008","journal-title":"J. Bacteriol."},{"key":"ref54","doi-asserted-by":"publisher","first-page":"603","DOI":"10.1128\/CMR.00134-14","article-title":"Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management","volume":"28","author":"Tong","year":"2015","journal-title":"Clin. Microbiol. Rev."},{"key":"ref56","doi-asserted-by":"publisher","first-page":"1114","DOI":"10.1128\/JB.00954-15","article-title":"SrrAB modulates Staphylococcus aureus cell death through regulation of cidABC transcription","volume":"198","author":"Windham","year":"2016","journal-title":"J. Bacteriol."},{"key":"ref57","doi-asserted-by":"publisher","first-page":"458","DOI":"10.1111\/j.1365-2958.2006.05105.x","article-title":"Characterization of the Staphylococcus aureus CidR regulon: elucidation of a novel role for acetoin metabolism in cell death and lysis","volume":"60","author":"Yang","year":"2006","journal-title":"Mol. Microbiol."},{"key":"ref58","doi-asserted-by":"publisher","first-page":"603","DOI":"10.1016\/j.ijmm.2014.04.003","article-title":"Staphylococcus aureus glucose-induced biofilm accessory proteins, GbaAB, influence biofilm formation in a PIA-dependent manner","volume":"304","author":"You","year":"2014","journal-title":"Int. J. Med. Microbiol."},{"key":"ref59","doi-asserted-by":"publisher","first-page":"4819","DOI":"10.1021\/acs.biochem.7b00570","article-title":"Staphylococcus aureusCidC is a pyruvate:menaquinone oxidoreductase","volume":"56","author":"Zhang","year":"2017","journal-title":"Biochemistry"}],"container-title":["Frontiers in Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2022.914512\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,6,3]],"date-time":"2022-06-03T09:30:14Z","timestamp":1654248614000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2022.914512\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,6,3]]},"references-count":55,"alternative-id":["10.3389\/fmicb.2022.914512"],"URL":"https:\/\/doi.org\/10.3389\/fmicb.2022.914512","relation":{},"ISSN":["1664-302X"],"issn-type":[{"type":"electronic","value":"1664-302X"}],"subject":[],"published":{"date-parts":[[2022,6,3]]},"article-number":"914512"}}