{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T22:07:14Z","timestamp":1768514834353,"version":"3.49.0"},"posted":{"date-parts":[[2024,9,25]]},"group-title":"Microbiology","reference-count":46,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2024,9,25]],"date-time":"2024-09-25T00:00:00Z","timestamp":1727222400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,9,25]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  The electrophile methylglyoxal (MG) is produced by microorganisms and host cells through central metabolic pathways. MG is a highly reactive electrophile, so it must be rapidly detoxified to prevent damaging modifications to macromolecules.\n                  <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n                  , a pathogen of concern due to its ability develop multidrug resistance, causes many types of infections that have been associated with elevated MG levels, including cystic fibrosis (CF).\n                  <jats:italic>P. aeruginosa<\/jats:italic>\n                  isolates commonly have mutations that lead to LasR loss-of-function (LasR-) and we found that\n                  <jats:italic>lasR<\/jats:italic>\n                  mutations confer sensitivity to MG in multiple strain backgrounds. LasR-strains have increased activity of the CbrAB two-component system which represses Crc regulation of metabolism. Here, we show that higher CbrAB activity and low Crc activity renders cells sensitive to MG. We found that\n                  <jats:italic>P. aeruginosa<\/jats:italic>\n                  LasR-strains are more sensitive to MG and have lower intracellular reduced glutathione (GSH) compared to their LasR+ comparators. Consistent with published reports, mutants lacking\n                  <jats:italic>gloA3<\/jats:italic>\n                  , which encodes a MG-glyoxalase, and mutants lacking GSH biosynthesis enzymes (\n                  <jats:italic>gshA<\/jats:italic>\n                  or\n                  <jats:italic>gshB<\/jats:italic>\n                  ) were sensitive to MG. Exogenous GSH rescued MG sensitivity in LasR-strains and\n                  <jats:italic>gshA<\/jats:italic>\n                  or\n                  <jats:italic>gshB<\/jats:italic>\n                  mutants, but not in a\n                  <jats:italic>gloA3<\/jats:italic>\n                  mutant strain. We propose that low GSH levels in LasR-strains contribute to increased sensitivity to MG and H\n                  <jats:sub>2<\/jats:sub>\n                  O\n                  <jats:sub>2<\/jats:sub>\n                  .\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Significance<\/jats:title>\n                  <jats:p>\n                    Methylglyoxal is a highly reactive metabolite that is detected in various disease states, including those where\n                    <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n                    is present and MG resistance requires the glutathione-dependent glyoxalase enzyme GloA3 enzyme. This study reveals that\n                    <jats:italic>P<\/jats:italic>\n                    .\n                    <jats:italic>aeruginosa<\/jats:italic>\n                    strains with LasR mutations, which are commonly found in clinical isolates, are more sensitive to methylglyoxal (MG) and hydrogen peroxide due to lower intracellular glutathione levels and high activity of the CbrAB-Crc regulatory pathway. This could be significant for understanding the selective pressures that drive\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    evolution in infection sites, as well as a better understanding of LasR-strain metabolism in infections such as those associated with cystic fibrosis.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2024.09.25.615034","type":"posted-content","created":{"date-parts":[[2024,9,26]],"date-time":"2024-09-26T00:15:12Z","timestamp":1727309712000},"source":"Crossref","is-referenced-by-count":0,"title":["<i>Pseudomonas aeruginosa<\/i>\n                  LasR-deficient mutants have increased methylglyoxal and hydrogen peroxide sensitivity due to low intracellular glutathione"],"prefix":"10.64898","author":[{"given":"Marina","family":"Ruzic","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana I.","family":"Altamirano Hefferan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Amy","family":"Conaway","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6366-2971","authenticated-orcid":false,"given":"Deborah A.","family":"Hogan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2024100104451057000_2024.09.25.615034v1.1","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00001.2019"},{"key":"2024100104451057000_2024.09.25.615034v1.2","doi-asserted-by":"publisher","DOI":"10.1042\/BJ20121743"},{"key":"2024100104451057000_2024.09.25.615034v1.3","doi-asserted-by":"publisher","DOI":"10.1042\/bj3050999"},{"key":"2024100104451057000_2024.09.25.615034v1.4","doi-asserted-by":"crossref","first-page":"471","DOI":"10.3390\/cryst9090471","article-title":"Crystal Structure of NADPH-Dependent Methylglyoxal Reductase Gre2 from Candida Albicans","volume":"9","year":"2019","journal-title":"Crystals"},{"key":"2024100104451057000_2024.09.25.615034v1.5","doi-asserted-by":"publisher","DOI":"10.1042\/bj2690001"},{"key":"2024100104451057000_2024.09.25.615034v1.6","doi-asserted-by":"publisher","DOI":"10.1007\/BF00808119"},{"key":"2024100104451057000_2024.09.25.615034v1.7","doi-asserted-by":"publisher","DOI":"10.2337\/diabetes.48.1.198"},{"key":"2024100104451057000_2024.09.25.615034v1.8","doi-asserted-by":"publisher","DOI":"10.1007\/s00125-013-2919-8"},{"key":"2024100104451057000_2024.09.25.615034v1.9","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1016\/j.semcancer.2017.05.010","article-title":"Methylglyoxal-derived stress: An emerging biological factor involved in the onset and progression of cancer","volume":"49","year":"2018","journal-title":"Semin Cancer Biol"},{"key":"2024100104451057000_2024.09.25.615034v1.10","doi-asserted-by":"publisher","DOI":"10.1016\/j.mad.2011.10.007"},{"key":"2024100104451057000_2024.09.25.615034v1.11","doi-asserted-by":"crossref","unstructured":"Pariano M , Costantini C , Santarelli I , Puccetti M , Giovagnoli S , Talesa VN , Romani L , Antognelli C. 2021. Defective Glyoxalase 1 Contributes to Pathogenic Inflammation in Cystic Fibrosis. Vaccines (Basel) 9.","DOI":"10.3390\/vaccines9111311"},{"key":"2024100104451057000_2024.09.25.615034v1.12","doi-asserted-by":"publisher","DOI":"10.1515\/BC.2009.033"},{"key":"2024100104451057000_2024.09.25.615034v1.13","doi-asserted-by":"crossref","unstructured":"Mey JT , Haus JM . 2018. Dicarbonyl Stress and Glyoxalase-1 in Skeletal Muscle: Implications for Insulin Resistance and Type 2 Diabetes. Frontiers in Cardiovascular Medicine 5.","DOI":"10.3389\/fcvm.2018.00117"},{"key":"2024100104451057000_2024.09.25.615034v1.14","doi-asserted-by":"publisher","DOI":"10.1152\/jappl.1993.75.6.2419"},{"key":"2024100104451057000_2024.09.25.615034v1.15","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbapap.2007.04.005"},{"key":"2024100104451057000_2024.09.25.615034v1.16","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbagen.2012.09.008"},{"key":"2024100104451057000_2024.09.25.615034v1.17","doi-asserted-by":"crossref","unstructured":"Michie KL , Dees JL , Fleming D , Moustafa DA , Goldberg JB , Rumbaugh KP , Whiteley M. 2020. Role of Pseudomonas aeruginosa Glutathione Biosynthesis in Lung and Soft Tissue Infection. Infect Immun 88.","DOI":"10.1128\/IAI.00116-20"},{"key":"2024100104451057000_2024.09.25.615034v1.18","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-020-58062-y"},{"key":"2024100104451057000_2024.09.25.615034v1.19","doi-asserted-by":"crossref","first-page":"164","DOI":"10.3389\/fcimb.2019.00164","article-title":"Glutathione Activates Type III Secretion System Through Vfr in Pseudomonas aeruginosa","volume":"9","year":"2019","journal-title":"Front Cell Infect Microbiol"},{"key":"2024100104451057000_2024.09.25.615034v1.20","doi-asserted-by":"publisher","DOI":"10.1007\/s11427-009-0074-8"},{"key":"2024100104451057000_2024.09.25.615034v1.21","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0205815"},{"key":"2024100104451057000_2024.09.25.615034v1.22","doi-asserted-by":"crossref","unstructured":"Bouzo D , Cokcetin NN , Li L , Ballerin G , Bottomley AL , Lazenby J , Whitchurch CB , Paulsen IT , Hassan KA , Harry EJ . 2020. Characterizing the Mechanism of Action of an Ancient Antimicrobial, Manuka Honey, against Pseudomonas aeruginosa Using Modern Transcriptomics. mSystems 5.","DOI":"10.1128\/msystems.00106-20"},{"key":"2024100104451057000_2024.09.25.615034v1.23","first-page":"180","article-title":"Effect of methylglyoxal on multidrug-resistant Pseudomonas aeruginosa","volume":"5","year":"2014","journal-title":"Front Microbiol"},{"key":"2024100104451057000_2024.09.25.615034v1.24","doi-asserted-by":"publisher","DOI":"10.1128\/JB.185.7.2066-2079.2003"},{"key":"2024100104451057000_2024.09.25.615034v1.25","doi-asserted-by":"crossref","unstructured":"Mould DL , Stevanovic M , Ashare A , Schultz D , Hogan DA . 2022. Metabolic basis for the evolution of a common pathogenic Pseudomonas aeruginosa variant. Elife 11.","DOI":"10.7554\/eLife.76555"},{"key":"2024100104451057000_2024.09.25.615034v1.26","doi-asserted-by":"publisher","DOI":"10.1111\/1462-2920.12499"},{"key":"2024100104451057000_2024.09.25.615034v1.27","doi-asserted-by":"crossref","first-page":"898","DOI":"10.1111\/1462-2920.14471","article-title":"The global regulator Crc orchestrates the metabolic robustness underlying oxidative stress resistance in Pseudomonas aeruginosa","volume":"21","year":"2019","journal-title":"Environmental Microbiology"},{"key":"2024100104451057000_2024.09.25.615034v1.28","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462-2920.2010.02292.x"},{"key":"2024100104451057000_2024.09.25.615034v1.29","doi-asserted-by":"publisher","DOI":"10.1016\/j.jcf.2008.09.006"},{"key":"2024100104451057000_2024.09.25.615034v1.30","doi-asserted-by":"publisher","DOI":"10.1038\/ismej.2014.98"},{"key":"2024100104451057000_2024.09.25.615034v1.31","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.1999.01672.x"},{"key":"2024100104451057000_2024.09.25.615034v1.32","doi-asserted-by":"publisher","DOI":"10.1128\/JB.182.16.4533-4544.2000"},{"key":"2024100104451057000_2024.09.25.615034v1.33","doi-asserted-by":"publisher","DOI":"10.1007\/s10541-005-0248-3"},{"key":"2024100104451057000_2024.09.25.615034v1.34","doi-asserted-by":"publisher","DOI":"10.1155\/2012\/736837"},{"key":"2024100104451057000_2024.09.25.615034v1.35","doi-asserted-by":"crossref","unstructured":"Zhang MM , Ong CL , Walker MJ , McEwan AG . 2016. Defence against methylglyoxal in Group A Streptococcus: a role for Glyoxylase I in bacterial virulence and survival in neutrophils? Pathog Dis 74.","DOI":"10.1093\/femspd\/ftv122"},{"key":"2024100104451057000_2024.09.25.615034v1.36","doi-asserted-by":"crossref","first-page":"e0108622","DOI":"10.1128\/mbio.01086-22","article-title":"High Glucose and Carbonyl Stress Impair HIF-1-Regulated Responses and the Control of Mycobacterium tuberculosis in Macrophages","volume":"13","year":"2022","journal-title":"mBio"},{"key":"2024100104451057000_2024.09.25.615034v1.37","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1111\/mmi.14604","article-title":"Mrr1 regulation of methylglyoxal catabolism and methylglyoxal-induced fluconazole resistance in Candida lusitaniae","volume":"115","year":"2021","journal-title":"Mol Microbiol"},{"key":"2024100104451057000_2024.09.25.615034v1.38","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1807698115"},{"key":"2024100104451057000_2024.09.25.615034v1.39","doi-asserted-by":"crossref","unstructured":"Juarez P , Jeannot K , Pl\u00e9siat P , Llanes C. 2017. Toxic Electrophiles Induce Expression of the Multidrug Efflux Pump MexEF-OprN in Pseudomonas aeruginosa through a Novel Transcriptional Regulator, CmrA. Antimicrob Agents Chemother 61.","DOI":"10.1128\/AAC.00585-17"},{"key":"2024100104451057000_2024.09.25.615034v1.40","doi-asserted-by":"crossref","unstructured":"Zhou H , Wang M , Smalley NE , Kostylev M , Schaefer AL , Greenberg EP , Dandekar AA , Xu F. 2019. Modulation of Pseudomonas aeruginosa Quorum Sensing by Glutathione. J Bacteriol 201.","DOI":"10.1128\/JB.00685-18"},{"key":"2024100104451057000_2024.09.25.615034v1.41","doi-asserted-by":"crossref","first-page":"e0067622","DOI":"10.1128\/mbio.00676-22","article-title":"Streptococcus pyogenes Hijacks Host Glutathione for Growth and Innate Immune Evasion","volume":"13","year":"2022","journal-title":"mBio"},{"key":"2024100104451057000_2024.09.25.615034v1.42","doi-asserted-by":"crossref","first-page":"102012","DOI":"10.1016\/j.redox.2021.102012","article-title":"New roles for glutathione: Modulators of bacterial virulence and pathogenesis","volume":"44","year":"2021","journal-title":"Redox Biol"},{"key":"2024100104451057000_2024.09.25.615034v1.43","doi-asserted-by":"crossref","unstructured":"Van Laar TA , Esani S , Birges TJ , Hazen B , Thomas JM , Rawat M. 2018. Pseudomonas aeruginosa gshA Mutant Is Defective in Biofilm Formation, Swarming, and Pyocyanin Production. mSphere 3.","DOI":"10.1128\/mSphere.00155-18"},{"key":"2024100104451057000_2024.09.25.615034v1.44","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M113.505784"},{"key":"2024100104451057000_2024.09.25.615034v1.45","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2018.06.064"},{"key":"2024100104451057000_2024.09.25.615034v1.46","doi-asserted-by":"crossref","unstructured":"Kim HJ , Lee KY , Kwon AR , Lee BJ . 2017. Structural and functional studies of SAV0551 from Staphylococcus aureus as a chaperone and glyoxalase III. Biosci Rep 37.","DOI":"10.1042\/BSR20171106"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2024.09.25.615034","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T07:08:13Z","timestamp":1768460893000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2024.09.25.615034"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,9,25]]},"references-count":46,"URL":"https:\/\/doi.org\/10.1101\/2024.09.25.615034","relation":{},"subject":[],"published":{"date-parts":[[2024,9,25]]},"subtype":"preprint"}}