{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T23:36:57Z","timestamp":1792798617850,"version":"4.5.0"},"reference-count":69,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2024,1,11]],"date-time":"2024-01-11T00:00:00Z","timestamp":1704931200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2024,1,11]],"date-time":"2024-01-11T00:00:00Z","timestamp":1704931200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"name":"HHS | National Institutes of Health","award":["DP2GM146456-01"],"award-info":[{"award-number":["DP2GM146456-01"]}]},{"name":"HHS | NIH | National Institute of Dental and Craniofacial Research","award":["F30DE032598, T90DE021989-11"],"award-info":[{"award-number":["F30DE032598, T90DE021989-11"]}],"id":[{"id":"https:\/\/ror.org\/004a2wv92","id-type":"ROR","asserted-by":"crossref"}]},{"name":"HHS | National Institutes of Health","award":["GM117061"],"award-info":[{"award-number":["GM117061"]}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2024,1,11]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    The ubiquitous opportunistic pathogen\n                    <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n                    is highly adaptive and refractory to several different classes of antibiotics. However, we found in this study that stationary-phase\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    cultures exhibit greater sensitivity to the fluoroquinolone Levofloxacin (Levo) than other bactericidal antibiotics, including an aminoglycoside (Tobramycin) and \u03b2-lactam (Aztreonam). To understand the basis of this sensitivity, we conducted time-lapse fluorescence microscopy experiments of cells during Levo treatment. We discovered that stationary-phase\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    cells die rapidly during treatment and undergo heterogeneous morphological changes, including explosive lysis, filamentation, and gradual loss of membrane integrity as evidenced by propidium iodide uptake. These morphologies are reminiscent of how the model organism\n                    <jats:italic>Escherichia coli<\/jats:italic>\n                    appears when recovering from fluoroquinolone treatment, a period when activation of the DNA damage-induced SOS response is crucial. Accordingly, we monitored the morphologies and survival of\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    \u0394\n                    <jats:italic>recA<\/jats:italic>\n                    mutants and found that the SOS response is not involved in\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    Levo sensitivity like it is for\n                    <jats:italic>E. coli<\/jats:italic>\n                    . We hypothesized that Levo sensitivity may be due to\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    maintaining active metabolism in stationary phase. We determined that stationary-phase\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    cells transcribe, maintain reductase activity, and accumulate reactive metabolic species which contribute to Levo-mediated death. By elucidating how\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    cells sustain metabolic activity during the stationary phase, we can design strategies to sensitize these persistent subpopulations to Levo and maintain the efficacy of this clinically important fluoroquinolone antibiotic.\n                  <\/jats:p>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      The bacterial pathogen\n                      <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n                      is responsible for a variety of chronic human infections. Even in the absence of identifiable resistance mutations, this pathogen can tolerate lethal antibiotic doses through phenotypic strategies like biofilm formation and metabolic quiescence. In this study, we determined that\n                      <jats:italic>P. aeruginosa<\/jats:italic>\n                      maintains greater metabolic activity in the stationary phase compared to the model organism,\n                      <jats:italic>Escherichia coli<\/jats:italic>\n                      , which has traditionally been used to study fluoroquinolone antibiotic tolerance. We demonstrate that hallmarks of\n                      <jats:italic>E. coli<\/jats:italic>\n                      fluoroquinolone tolerance are not conserved in\n                      <jats:italic>P. aeruginosa<\/jats:italic>\n                      , including the timing of cell death and necessity of the SOS DNA damage response for survival. The heightened sensitivity of stationary-phase\n                      <jats:italic>P. aeruginosa<\/jats:italic>\n                      to fluoroquinolones is attributed to maintained transcriptional and reductase activity. Our data suggest that perturbations that suppress transcription and respiration in\n                      <jats:italic>P. aeruginosa<\/jats:italic>\n                      may actually protect the pathogen against this important class of antibiotics.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.03567-23","type":"journal-article","created":{"date-parts":[[2023,12,11]],"date-time":"2023-12-11T09:00:34Z","timestamp":1702285234000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":10,"title":["Metabolic and transcriptional activities underlie stationary-phase\n                    <i>Pseudomonas aeruginosa<\/i>\n                    sensitivity to Levofloxacin"],"prefix":"10.1128","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5014-6161","authenticated-orcid":false,"given":"Patricia J.","family":"Hare","sequence":"first","affiliation":[{"name":"Department of Molecular Biology &amp; Biophysics, UConn Health , Farmington, Connecticut, USA"},{"name":"School of Dental Medicine, UConn Health , Farmington, Connecticut, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juliet R.","family":"Gonzalez","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology &amp; Biophysics, UConn Health , Farmington, Connecticut, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryan M.","family":"Quelle","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology &amp; Biophysics, UConn Health , Farmington, Connecticut, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi I.","family":"Wu","sequence":"additional","affiliation":[{"name":"Richard D. Berlin Center for Cell Analysis and Modeling, UConn Health , Farmington, Connecticut, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6638-2959","authenticated-orcid":true,"given":"Wendy W. K.","family":"Mok","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology &amp; Biophysics, UConn Health , Farmington, Connecticut, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01651-09"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1099\/jmm.0.030932-0"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro.2016.34"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-019-0196-3"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bioflm.2021.100056"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2015.05.011"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.3001194"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2016.03.035"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.aaf4762"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1804525115"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.47.1.317-323.2003"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-28141-x"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature10069"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2014.00070"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1211037"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.02061-12"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-08733-w"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2016.12.015"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.35.9.1824"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.183.23.6746-6751.2001"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.2009.01717.x"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro.2016.107"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-020-00477-5"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pgen.1003144"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00731-15"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1099390"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.aav9462"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00505-07"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-019-03377-x"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.42.7.1641"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.28.2.331"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01231-09"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.5507\/bp.2016.057"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.48.7.2659-2664.2004"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2022.1023523"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1097\/QCO.0000000000000677"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-09058-4"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.02230-14"},{"key":"e_1_3_4_40_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.163.2.568-572.1985"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.1038\/ismej.2016.79"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.180.15.3946-3953.1998"},{"key":"e_1_3_4_43_2","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics11030325"},{"key":"e_1_3_4_44_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.02107-16"},{"key":"e_1_3_4_45_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00807-06"},{"key":"e_1_3_4_46_2","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms11220"},{"key":"e_1_3_4_47_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01889-14"},{"key":"e_1_3_4_48_2","doi-asserted-by":"publisher","DOI":"10.1128\/mmbr.61.3.377-392.1997"},{"key":"e_1_3_4_49_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1804218115"},{"key":"e_1_3_4_50_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00085-11"},{"key":"e_1_3_4_51_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.01166-10"},{"key":"e_1_3_4_52_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-2854-5"},{"key":"e_1_3_4_53_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2007.06.049"},{"key":"e_1_3_4_54_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1401876111"},{"key":"e_1_3_4_55_2","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.02312-22"},{"key":"e_1_3_4_56_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gky1143"},{"key":"e_1_3_4_57_2","doi-asserted-by":"publisher","DOI":"10.1128\/aac.01639-22"},{"key":"e_1_3_4_58_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2212340119"},{"key":"e_1_3_4_59_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro3316"},{"key":"e_1_3_4_60_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.174.2.345-348.1992"},{"key":"e_1_3_4_61_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462-2920.2012.02759.x"},{"key":"e_1_3_4_62_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2019.02603"},{"key":"e_1_3_4_63_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/8.4.249"},{"key":"e_1_3_4_64_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1901730116"},{"key":"e_1_3_4_65_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cub.2021.02.040"},{"key":"e_1_3_4_66_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2015.115"},{"key":"e_1_3_4_67_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.100.1.215-219.1969"},{"key":"e_1_3_4_68_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymben.2017.03.006"},{"key":"e_1_3_4_69_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.173.15.4700-4706.1991"},{"key":"e_1_3_4_70_2","volume-title":"Performance standards for antimicrobial susceptibility testing","author":"Clinical and Laboratory Standards Institute","year":"2021","unstructured":"Clinical and Laboratory Standards Institute . 2021. Performance standards for antimicrobial susceptibility testing. 31st ed. Wayne, PA.","edition":"31"}],"container-title":["Microbiology Spectrum"],"original-title":[],"contributor":[{"given":"Gregory","family":"Wiedman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"reviewer"}]}],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.03567-23","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.03567-23","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,11]],"date-time":"2024-01-11T09:17:14Z","timestamp":1704964634000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/spectrum.03567-23"}},"subtitle":[],"editor":[{"given":"Minsu","family":"Kim","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2024,1,11]]},"references-count":69,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2024,1,11]]}},"alternative-id":["10.1128\/spectrum.03567-23"],"URL":"https:\/\/doi.org\/10.1128\/spectrum.03567-23","relation":{},"ISSN":["2165-0497"],"issn-type":[{"value":"2165-0497","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,1,11]]},"assertion":[{"value":"2023-10-05","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2023-11-16","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2023-12-11","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e03567-23"}}