{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T06:55:27Z","timestamp":1790751327401,"version":"4.1.0"},"reference-count":116,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T00:00:00Z","timestamp":1790726400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Marie Sk\u0142odowska-Curie"},{"name":"Italian Ministry of University and Research"},{"name":"Italian Ministry of University and Research"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:p>Gram-negative bacteria are a major cause of antibiotic-resistant infections due to their highly impermeable cell envelope which contains lipopolysaccharide (LPS) and prevents antibiotics from entering the cell. LPS biogenesis relies on the activity of the essential LPS transport machinery (Lpt), which represents an attractive and validated target for antibiotic development. The antimicrobial peptide thanatin impairs LPS transport by disrupting the assembly of the Lpt protein machinery, resulting in cell death. However, thanatin is not a suitable drug candidate due to its poor drug-like properties and the rapid emergence of resistance. Therefore, understanding the mechanisms underlying cell resistance and adaptation to thanatin is crucial for developing novel antimicrobial strategies. Here we performed RNA-seq\u2013based kinetic transcriptomic analysis to investigate how bacterial cells respond to thanatin over time followed by a functional validation. We found that thanatin treatment induces a rapid activation of the Cpx envelope stress response system, followed by activation of Rcs phosphorelay and downregulation of the FlhDC regulon. Cpx regulated genes showed enrichment for pathways involved in cationic peptide resistance, stress regulation, protein folding, and peptidoglycan remodelling. Rcs upregulated genes were associated with capsule synthesis and biofilm formation. Finally, late response to thanatin suggests the involvement of phospholipid metabolism modulation and trafficking as a strategy to restore OM asymmetry. Overall, our results delineate the adaptive mechanisms employed by bacterial cells to counteract thanatin-induced stress through an interplay of multiple regulatory pathways controlling envelope integrity, stress responses, motility and biofilm formation.<\/jats:p>","DOI":"10.3389\/fmicb.2026.1920608","type":"journal-article","created":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T06:00:46Z","timestamp":1790748046000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Time-resolved transcriptomic analysis reveals envelope stress\u2013mediated adaptation of Escherichia coli to the LPS transport inhibitor thanatin"],"prefix":"10.3389","volume":"17","author":[{"given":"Renu","family":"Bisht","sequence":"first","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences \u201cRodolfo Paoletti\u201d, University of Milano","place":["Milano, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elisabete C. C. M.","family":"Moura","sequence":"additional","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences \u201cRodolfo Paoletti\u201d, University of Milano","place":["Milano, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julien","family":"Prados","sequence":"additional","affiliation":[{"name":"Bioinformatics Support Platform, Faculty of Medicine, University of Geneva","place":["Geneva, Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessandra","family":"Romanelli","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, University of Milano","place":["Milano, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessandra M.","family":"Martorana","sequence":"additional","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences \u201cRodolfo Paoletti\u201d, University of Milano","place":["Milano, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pierre D.","family":"Charlesworth","sequence":"additional","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences \u201cRodolfo Paoletti\u201d, University of Milano","place":["Milano, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M\u00e9lanie","family":"Roch","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Medicine, University Hospital and Medical School of Geneva","place":["Geneva, Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William L.","family":"Kelley","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Medicine, University Hospital and Medical School of Geneva","place":["Geneva, Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paola","family":"Sperandeo","sequence":"additional","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences \u201cRodolfo Paoletti\u201d, University of Milano","place":["Milano, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessandra","family":"Polissi","sequence":"additional","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences \u201cRodolfo Paoletti\u201d, University of Milano","place":["Milano, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,9,30]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"1996","DOI":"10.1099\/mic.0.000365","article-title":"A feed-forward loop between SroC and MgrR small RNAs modulates the expression of eptB and the susceptibility to polymyxin B in Salmonella Typhimurium","volume":"162","author":"Acu\u00f1a","year":"2016","journal-title":"Microbiology (Reading)"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"143","DOI":"10.1128\/jb.176.1.143-148.1994","article-title":"Repressor mutations in the marRAB operon that activate oxidative stress genes and multiple antibiotic resistance in Escherichia coli","volume":"176","author":"Ariza","year":"1994","journal-title":"J. Bacteriol."},{"key":"ref3","doi-asserted-by":"publisher","first-page":"7770","DOI":"10.1128\/AEM.02593-13","article-title":"Induction of the Cpx envelope stress pathway contributes to Escherichia coli tolerance to antimicrobial peptides","volume":"79","author":"Audrain","year":"2013","journal-title":"Appl. Environ. Microbiol."},{"key":"ref4","doi-asserted-by":"publisher","first-page":"0008","DOI":"10.1038\/msb4100050","article-title":"Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection","volume":"2","author":"Baba","year":"2006","journal-title":"Mol. Syst. Biol."},{"key":"ref5","doi-asserted-by":"publisher","first-page":"603","DOI":"10.1128\/jb.02449-14","article-title":"The Cpx envelope stress response modifies peptidoglycan cross-linking via the L,D-transpeptidase LdtD and the novel protein YgaU","volume":"197","author":"Bernal-Cabas","year":"2015","journal-title":"J. Bacteriol."},{"key":"ref6","doi-asserted-by":"publisher","first-page":"889","DOI":"10.3390\/pathogens13100889","article-title":"Breaking barriers: exploiting envelope biogenesis and stress responses to develop novel antimicrobial strategies in gram-negative Bacteria","volume":"13","author":"Bisht","year":"2024","journal-title":"Pathogens"},{"key":"ref7","doi-asserted-by":"publisher","first-page":"1453","DOI":"10.1126\/science.277.5331.1453","article-title":"The complete genome sequence of Escherichia coli K-12","volume":"277","author":"Blattner","year":"1997","journal-title":"Science"},{"key":"ref8","doi-asserted-by":"publisher","first-page":"971","DOI":"10.1046\/j.1365-2958.1998.00855.x","article-title":"Interplay between global regulators of Escherichia coli: effect of RpoS, Lrp and H-NS on transcription of the gene osmC","volume":"28","author":"Bouvier","year":"1998","journal-title":"Mol. Microbiol."},{"key":"ref9","doi-asserted-by":"publisher","first-page":"83","DOI":"10.1146\/annurev-micro-092412-155751","article-title":"The biology of the PmrA\/PmrB two-component system: the major regulator of lipopolysaccharide modifications","volume":"67","author":"Chen","year":"2013","journal-title":"Ann. Rev. Microbiol."},{"key":"ref10","doi-asserted-by":"publisher","first-page":"1300","DOI":"10.1016\/j.cell.2014.11.017","article-title":"Beta-lactam antibiotics induce a lethal malfunctioning of the bacterial Cell Wall synthesis machinery","volume":"159","author":"Cho","year":"2014","journal-title":"Cell"},{"key":"ref11","doi-asserted-by":"publisher","first-page":"640","DOI":"10.1128\/aac.02095-13","article-title":"Functional linkage between genes that regulate osmotic stress responses and multidrug resistance transporters: challenges and opportunities for antibiotic discovery","volume":"58","author":"Cohen","year":"2014","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref12","doi-asserted-by":"publisher","first-page":"191","DOI":"10.1146\/annurev-micro-041522-102901","article-title":"Periplasmic chaperones: outer membrane biogenesis and envelope stress","volume":"78","author":"Combs","year":"2024","journal-title":"Ann. Rev. Microbiol."},{"key":"ref13","doi-asserted-by":"publisher","first-page":"1183","DOI":"10.1101\/gad.11.9.1183","article-title":"The sigma(E) and the Cpx signal transduction systems control the synthesis of periplasmic protein-folding enzymes in Escherichia coli","volume":"11","author":"Danese","year":"1997","journal-title":"Genes Dev."},{"key":"ref14","doi-asserted-by":"publisher","first-page":"1522","DOI":"10.3390\/ijms22041522","article-title":"Thanatin: an emerging host defense antimicrobial peptide with multiple modes of action","volume":"22","author":"Dash","year":"2021","journal-title":"Int. J. Mol. Sci."},{"key":"ref15","doi-asserted-by":"publisher","first-page":"6640","DOI":"10.1073\/pnas.120163297","article-title":"One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products","volume":"97","author":"Datsenko","year":"2000","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref16","doi-asserted-by":"publisher","first-page":"e1002184","DOI":"10.1371\/journal.pmed.1002184","article-title":"Will 10 million people die a year due to antimicrobial resistance by 2050?","volume":"13","author":"de Kraker","year":"2016","journal-title":"PLoS Med."},{"key":"ref17","doi-asserted-by":"publisher","first-page":"461","DOI":"10.1128\/jb.01157-08","article-title":"The Crp-activated small noncoding regulatory RNA CyaR (RyeE) links nutritional status to group behavior","volume":"191","author":"De Lay","year":"2009","journal-title":"J. Bacteriol."},{"key":"ref18","doi-asserted-by":"publisher","first-page":"380","DOI":"10.3389\/fcimb.2019.00380","article-title":"A fly on the wall: how stress response systems can sense and respond to damage to peptidoglycan [review]","volume":"9","author":"Delhaye","year":"2019","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref19","doi-asserted-by":"publisher","first-page":"311","DOI":"10.1111\/j.1365-2958.1993.tb01122.x","article-title":"Regulation of transcription of genes required for fatty acid transport and unsatu rated fatty acid biosynthesis in Escherichia coli by FadR","volume":"7","author":"DiRusso","year":"1993","journal-title":"Mol. Microbiol."},{"key":"ref20","doi-asserted-by":"publisher","first-page":"579","DOI":"10.1016\/S0021-9258(19)77781-3","article-title":"A new spectrophotometric test for the detection of methylpentose","volume":"192","author":"Dische","year":"1951","journal-title":"J. Biol. Chem."},{"key":"ref21","doi-asserted-by":"publisher","first-page":"e2018329118","DOI":"10.1073\/pnas.2018329118","article-title":"Cardiolipin aids in lipopolysaccharide transport to the gram-negative outer membrane","volume":"118","author":"Douglass","year":"2021","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref22","doi-asserted-by":"publisher","first-page":"e0056726","DOI":"10.1128\/mbio.00567-26","article-title":"Lysophospholipid signaling coordinates outer membrane homeostasis in Escherichia coli","volume":"17","author":"Enverso","year":"2026","journal-title":"MBio"},{"key":"ref23","doi-asserted-by":"publisher","first-page":"1573","DOI":"10.1111\/jam.15314","article-title":"Antimicrobial peptides (AMPs): a promising class of antimicrobial compounds","volume":"132","author":"Erdem Buyukkiraz","year":"2022","journal-title":"J. Appl. Microbiol."},{"key":"ref24","doi-asserted-by":"publisher","first-page":"30960","DOI":"10.1038\/srep30960","article-title":"Reduced protein synthesis Fidelity inhibits Flagellar biosynthesis and motility","volume":"6","author":"Fan","year":"2016","journal-title":"Sci. Rep."},{"key":"ref25","doi-asserted-by":"publisher","first-page":"1221","DOI":"10.1073\/pnas.93.3.1221","article-title":"Structure-activity analysis of thanatin, a 21-residue inducible insect defense peptide with sequence homology to frog skin antimicrobial peptides","volume":"93","author":"Fehlbaum","year":"1996","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref26","doi-asserted-by":"publisher","first-page":"1070","DOI":"10.1099\/mic.0.2006\/002907-0","article-title":"The yjbEFGH locus in Escherichia coli K-12 is an operon encoding proteins involved in exopolysaccharide production","volume":"153","author":"Ferrieres","year":"2007","journal-title":"Microbiology (Reading)"},{"key":"ref27","doi-asserted-by":"publisher","first-page":"187","DOI":"10.1038\/s41589-020-00694-2","article-title":"Dynamics of an LPS translocon induced by substrate and an antimicrobial peptide","volume":"17","author":"Fiorentino","year":"2021","journal-title":"Nat. Chem. Biol."},{"key":"ref28","doi-asserted-by":"publisher","first-page":"3873","DOI":"10.1128\/jb.187.11.3873-3877.2005","article-title":"Osmotic regulation of the Escherichia coli bdm (biofilm-dependent modulation) gene by the RcsCDB his-asp phosphorelay","volume":"187","author":"Francez-Charlot","year":"2005","journal-title":"J. Bacteriol."},{"key":"ref29","doi-asserted-by":"publisher","first-page":"823","DOI":"10.1046\/j.1365-2958.2003.03601.x","article-title":"RcsCDB his-asp phosphorelay system negatively regulates the flhDC operon in Escherichia coli","volume":"49","author":"Francez-Charlot","year":"2003","journal-title":"Mol. Microbiol."},{"key":"ref30","doi-asserted-by":"publisher","first-page":"2486","DOI":"10.1073\/pnas.1015617108","article-title":"The complex that inserts lipopolysaccharide into the bacterial outer membrane forms a two-protein plug-and-barrel","volume":"108","author":"Freinkman","year":"2011","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref31","doi-asserted-by":"publisher","first-page":"829","DOI":"10.1111\/j.1365-2958.2007.05947.x","article-title":"Regulation of fatty acid metabolism in bacteria","volume":"66","author":"Fujita","year":"2007","journal-title":"Mol. Microbiol."},{"key":"ref32","doi-asserted-by":"publisher","first-page":"1226","DOI":"10.1111\/febs.12701","article-title":"DegP primarily functions as a protease for the biogenesis of \u03b2-barrel outer membrane proteins in the gram-negative bacterium Escherichia coli","volume":"281","author":"Ge","year":"2014","journal-title":"FEBS J."},{"key":"ref33","doi-asserted-by":"publisher","first-page":"115446","DOI":"10.1016\/j.celrep.2025.115446","article-title":"Optimal functioning of the Lpt bridge depends on a ternary complex between the lipocalin YedD and the LptDE translocon","volume":"44","author":"Gennaris","year":"2025","journal-title":"Cell Rep."},{"key":"ref34","doi-asserted-by":"publisher","first-page":"14","DOI":"10.1186\/1741-7007-5-14","article-title":"A novel role for RecA under non-stress: promotion of swarming motility in Escherichia coli K-12","volume":"5","author":"G\u00f3mez-G\u00f3mez","year":"2007","journal-title":"BMC Biol."},{"key":"ref35","doi-asserted-by":"publisher","first-page":"e00312","DOI":"10.1128\/mBio.00312-16","article-title":"The CpxQ sRNA negatively regulates Skp to prevent mistargeting of beta-barrel outer membrane proteins into the cytoplasmic membrane","volume":"7","author":"Grabowicz","year":"2016","journal-title":"mBio"},{"key":"ref36","doi-asserted-by":"publisher","first-page":"232","DOI":"10.1016\/j.tibs.2016.10.002","article-title":"Envelope stress responses: an interconnected safety net","volume":"42","author":"Grabowicz","year":"2017","journal-title":"Trends Biochem. Sci."},{"key":"ref37","doi-asserted-by":"publisher","first-page":"81","DOI":"10.1111\/j.1574-6968.2012.02528.x","article-title":"Resistance to antimicrobial peptides in gram-negative bacteria","volume":"330","author":"Gruenheid","year":"2012","journal-title":"FEMS Microbiol. Lett."},{"key":"ref38","doi-asserted-by":"publisher","first-page":"849","DOI":"10.1111\/1574-6976.12018","article-title":"Regulation of flagellar motility during biofilm formation","volume":"37","author":"Guttenplan","year":"2013","journal-title":"FEMS Microbiol. Rev."},{"key":"ref39","doi-asserted-by":"publisher","first-page":"1551","DOI":"10.1038\/nbt1267","article-title":"Antimicrobial and host-defense peptides as new anti-infective therapeutic strategies","volume":"24","author":"Hancock","year":"2006","journal-title":"Nat. Biotechnol."},{"key":"ref40","doi-asserted-by":"publisher","first-page":"31029","DOI":"10.1074\/jbc.M114.603944","article-title":"The essential protein for bacterial flagella formation FlgJ functions as a \u03b2-N-acetylglucosaminidase","volume":"289","author":"Herlihey","year":"2014","journal-title":"J. Biol. Chem."},{"key":"ref41","doi-asserted-by":"publisher","first-page":"313","DOI":"10.3389\/fcimb.2019.00313","article-title":"Maintaining integrity under stress: envelope stress response regulation of pathogenesis in gram-negative Bacteria","volume":"9","author":"Hews","year":"2019","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref42","doi-asserted-by":"publisher","first-page":"e00638","DOI":"10.1128\/mSphere.00638-19","article-title":"Role of yoaE gene regulated by CpxR in the survival of Salmonella enterica serovar Enteritidis in antibacterial egg white","volume":"5","author":"Huang","year":"2020","journal-title":"mSphere"},{"key":"ref43","doi-asserted-by":"publisher","first-page":"219","DOI":"10.1007\/s00203-007-0311-0","article-title":"Induction of the yjbEFGH operon is regulated by growth rate and oxygen concentration","volume":"189","author":"Ionescu","year":"2008","journal-title":"Arch. Microbiol."},{"key":"ref44","doi-asserted-by":"publisher","first-page":"10574","DOI":"10.1016\/s0021-9258(18)86985-x","article-title":"Transcription of osmB, a gene encoding an Escherichia coli lipoprotein, is regulated by dual signals. Osmotic stress and stationary phase","volume":"265","author":"Jung","year":"1990","journal-title":"J. Biol. Chem."},{"key":"ref45","doi-asserted-by":"publisher","first-page":"634","DOI":"10.1038\/nrmicro2405","article-title":"A field guide to bacterial swarming motility","volume":"8","author":"Kearns","year":"2010","journal-title":"Nat. Rev. Microbiol."},{"key":"ref46","doi-asserted-by":"publisher","first-page":"e00519","DOI":"10.1128\/jb.00519-17","article-title":"A small periplasmic protein with a hydrophobic C-terminal residue enhances DegP proteolysis as a suicide activator","volume":"200","author":"Kim","year":"2018","journal-title":"J. Bacteriol."},{"key":"ref47","doi-asserted-by":"publisher","first-page":"15369","DOI":"10.1074\/jbc.M900490200","article-title":"Escherichia coli K-12 suppressor-free mutants lacking early glycosyltransferases and late acyltransferases: minimal lipopolysaccharide structure and induction of envelope stress response","volume":"284","author":"Klein","year":"2009","journal-title":"J. Biol. Chem."},{"key":"ref48","doi-asserted-by":"publisher","first-page":"100","DOI":"10.1111\/j.1365-2958.2007.05893.x","article-title":"Escherichia coli phage-shock protein a (PspA) binds to membrane phospholipids and repairs proton leakage of the damaged membranes","volume":"66","author":"Kobayashi","year":"2007","journal-title":"Mol. Microbiol."},{"key":"ref49","doi-asserted-by":"publisher","first-page":"1467726","DOI":"10.3389\/fmolb.2024.1467726","article-title":"Identification and validation of reference genes for quantitative gene expression analysis under 409 and 415 nm antimicrobial blue light treatment","volume":"11","author":"Kruszewska-Naczk","year":"2024","journal-title":"Front. Mol. Biosci."},{"key":"ref50","doi-asserted-by":"publisher","first-page":"2524","DOI":"10.1128\/jb.00320-16","article-title":"Evidence for Escherichia coli Diguanylate cyclase DgcZ interlinking surface sensing and adhesion via multiple regulatory routes","volume":"198","author":"Lacanna","year":"2016","journal-title":"J. Bacteriol."},{"key":"ref51","doi-asserted-by":"publisher","first-page":"473","DOI":"10.1111\/mmi.14168","article-title":"GcvB small RNA uses two distinct seed regions to regulate an extensive targetome","volume":"111","author":"Lalaouna","year":"2019","journal-title":"Mol. Microbiol."},{"key":"ref52","doi-asserted-by":"publisher","first-page":"837","DOI":"10.1126\/science.1235358","article-title":"Dual molecular signals mediate the bacterial response to outer-membrane stress","volume":"340","author":"Lima","year":"2013","journal-title":"Science"},{"key":"ref53","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1038\/s44259-024-00046-3","article-title":"Mechanisms of antimicrobial resistance in biofilms","volume":"2","author":"Liu","year":"2024","journal-title":"NPJ Antimicrob. Resist."},{"key":"ref54","doi-asserted-by":"publisher","first-page":"3877","DOI":"10.1099\/mic.0.2008\/022301-0","article-title":"Capsule polysaccharide is a bacterial decoy for antimicrobial peptides","volume":"154","author":"Llobet","year":"2008","journal-title":"Microbiology (Reading)"},{"key":"ref55","doi-asserted-by":"publisher","first-page":"55707","DOI":"10.1074\/jbc.M408994200","article-title":"Identification of a new member of the phage shock protein response in Escherichia coli, the phage shock protein G (PspG)","volume":"279","author":"Lloyd","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref56","doi-asserted-by":"publisher","first-page":"3162","DOI":"10.1128\/jb.00502-16","article-title":"Transcriptional responses of Escherichia coli to a small-molecule inhibitor of LolCDE, an essential component of the lipoprotein transport pathway","volume":"198","author":"Lorenz","year":"2016","journal-title":"J. Bacteriol."},{"key":"ref57","doi-asserted-by":"publisher","first-page":"e2110055118","DOI":"10.1073\/pnas.2110055118","article-title":"ATP disrupts lipid-binding equilibrium to drive retrograde transport critical for bacterial outer membrane asymmetry","volume":"118","author":"Low","year":"2021","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref58","doi-asserted-by":"publisher","first-page":"5098","DOI":"10.1021\/acs.chemrev.0c00587","article-title":"Assembly and maintenance of lipids at the bacterial outer membrane","volume":"121","author":"Lundstedt","year":"2021","journal-title":"Chem. Rev."},{"key":"ref59","doi-asserted-by":"publisher","first-page":"489","DOI":"10.1016\/0966-842x(94)90654-8","article-title":"Efflux pumps and drug resistance in gram-negative bacteria","volume":"2","author":"Ma","year":"1994","journal-title":"Trends Microbiol."},{"key":"ref60","doi-asserted-by":"publisher","first-page":"3517","DOI":"10.1038\/s41467-019-11503-3","article-title":"The antimicrobial peptide thanatin disrupts the bacterial outer membrane and inactivates the NDM-1 metallo-beta-lactamase","volume":"10","author":"Ma","year":"2019","journal-title":"Nat. Commun."},{"key":"ref61","doi-asserted-by":"publisher","first-page":"3927","DOI":"10.1128\/JB.00084-07","article-title":"Identification of the L,D-transpeptidases responsible for attachment of the Braun lipoprotein to Escherichia coli peptidoglycan","volume":"189","author":"Magnet","year":"2007","journal-title":"J. Bacteriol."},{"key":"ref62","doi-asserted-by":"publisher","first-page":"4782","DOI":"10.1128\/JB.00025-08","article-title":"Identification of the L,D-transpeptidases for peptidoglycan cross-linking in Escherichia coli","volume":"190","author":"Magnet","year":"2008","journal-title":"J. Bacteriol."},{"key":"ref63","doi-asserted-by":"publisher","first-page":"8009","DOI":"10.1073\/pnas.0903229106","article-title":"An ABC transport system that maintains lipid asymmetry in the gram-negative outer membrane","volume":"106","author":"Malinverni","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref64","doi-asserted-by":"publisher","first-page":"15174","DOI":"10.3390\/ijms242015174","article-title":"Suppressors of lapC mutation identify new regulators of LpxC, which mediates the first committed step in lipopolysaccharide biosynthesis","volume":"24","author":"Maniyeri","year":"2023","journal-title":"Int. J. Mol. Sci."},{"key":"ref65","doi-asserted-by":"publisher","first-page":"e100941","DOI":"10.1371\/journal.pone.0100941","article-title":"Dissecting Escherichia coli outer membrane biogenesis using differential proteomics","volume":"9","author":"Martorana","year":"2014","journal-title":"PLoS One"},{"key":"ref66","doi-asserted-by":"publisher","first-page":"758228","DOI":"10.3389\/fmolb.2021.758228","article-title":"Degradation of components of the Lpt transenvelope machinery reveals LPS-dependent Lpt complex stability in Escherichia coli","volume":"8","author":"Martorana","year":"2021","journal-title":"Front. Mol. Biosci."},{"key":"ref67","doi-asserted-by":"publisher","first-page":"e00379","DOI":"10.1128\/mBio.00379-18","article-title":"The Escherichia coli phospholipase PldA regulates outer membrane homeostasis via lipid signaling","volume":"9","author":"May","year":"2018","journal-title":"MBio"},{"key":"ref68","doi-asserted-by":"publisher","first-page":"298","DOI":"10.1128\/iai.00800-13","article-title":"Response of extraintestinal pathogenic Escherichia coli to human serum reveals a protective role for Rcs-regulated exopolysaccharide colanic acid","volume":"82","author":"Miajlovic","year":"2014","journal-title":"Infect. Immun."},{"key":"ref69","volume-title":"A Short Course in Bacterial Genetics: A Laboratory Manual and Handbook for","author":"Miller","year":"1992"},{"key":"ref70","doi-asserted-by":"publisher","first-page":"417","DOI":"10.1038\/s41579-019-0199-0","article-title":"Envelope stress responses: balancing damage repair and toxicity","volume":"17","author":"Mitchell","year":"2019","journal-title":"Nat. Rev. Microbiol."},{"key":"ref71","doi-asserted-by":"publisher","first-page":"e02729","DOI":"10.1128\/mBio.02729-18","article-title":"Peptidoglycan remodeling enables Escherichia coli to survive severe outer membrane assembly defect","volume":"10","author":"Mor\u00e8","year":"2019","journal-title":"mBio"},{"key":"ref72","doi-asserted-by":"publisher","first-page":"909","DOI":"10.3389\/fmicb.2020.00909","article-title":"Thanatin impairs lipopolysaccharide transport complex assembly by targeting LptC-LptA interaction and decreasing LptA stability","volume":"11","author":"Moura","year":"2020","journal-title":"Front. Microbiol."},{"key":"ref73","doi-asserted-by":"publisher","first-page":"629","DOI":"10.1016\/s0140-6736(21)02724-0","article-title":"Global burden of bacterial antimicrobial resistance in 2019: a systematic analysis","volume":"399","author":"Murray","year":"2022","journal-title":"Lancet"},{"key":"ref74","doi-asserted-by":"publisher","first-page":"4161","DOI":"10.1128\/jb.184.15.4161-4167.2002","article-title":"The putative response regulator BaeR stimulates multidrug resistance of Escherichia coli via a novel multidrug exporter system, MdtABC","volume":"184","author":"Nagakubo","year":"2002","journal-title":"J. Bacteriol."},{"key":"ref75","doi-asserted-by":"publisher","first-page":"435","DOI":"10.1111\/j.1365-2958.1992.tb01487.x","article-title":"Porins and specific channels of bacterial outer membranes","volume":"6","author":"Nikaido","year":"1992","journal-title":"Mol. Microbiol."},{"key":"ref76","doi-asserted-by":"publisher","first-page":"593","DOI":"10.1128\/MMBR.67.4.593-656.2003","article-title":"Molecular basis of bacterial outer membrane permeability revisited","volume":"67","author":"Nikaido","year":"2003","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref77","doi-asserted-by":"publisher","first-page":"4335","DOI":"10.3390\/molecules28114335","article-title":"Early molecular insights into thanatin analogues binding to A. baumannii LptA","volume":"28","author":"Oi","year":"2023","journal-title":"Molecules"},{"key":"ref78","doi-asserted-by":"publisher","first-page":"337","DOI":"10.1038\/nrmicro.2016.25","article-title":"Lipopolysaccharide transport and assembly at the outer membrane: the PEZ model","volume":"14","author":"Okuda","year":"2016","journal-title":"Nat. Rev. Microbiol."},{"key":"ref79","doi-asserted-by":"publisher","first-page":"1571","DOI":"10.1016\/j.chembiol.2024.07.002","article-title":"Insights into bacterial metabolism from small RNAs","volume":"31","author":"Papenfort","year":"2024","journal-title":"Cell. Chem. Biol."},{"key":"ref80","doi-asserted-by":"publisher","first-page":"2069","DOI":"10.1093\/jac\/dks196","article-title":"Bacterial stress responses as determinants of antimicrobial resistance","volume":"67","author":"Poole","year":"2012","journal-title":"J. Antimicrob. Chemother."},{"key":"ref81","doi-asserted-by":"publisher","first-page":"576","DOI":"10.1111\/j.1462-5822.2007.01111.x","article-title":"The salmonellae PhoQ sensor: mechanisms of detection of phagosome signals","volume":"10","author":"Prost","year":"2008","journal-title":"Cell. Microbiol."},{"key":"ref82","doi-asserted-by":"publisher","first-page":"2755","DOI":"10.1128\/JB.00105-13","article-title":"The Escherichia coli Cpx envelope stress response regulates genes of diverse function that impact antibiotic resistance and membrane integrity","volume":"195","author":"Raivio","year":"2013","journal-title":"J. Bacteriol."},{"key":"ref83","doi-asserted-by":"publisher","first-page":"176","DOI":"10.1186\/s12934-016-0577-5","article-title":"Genome-wide Escherichia coli stress response and improved tolerance towards industrially relevant chemicals","volume":"15","author":"Rau","year":"2016","journal-title":"Microb. Cell Factories"},{"key":"ref84","doi-asserted-by":"publisher","first-page":"5470","DOI":"10.1128\/jb.187.15.5470-5481.2005","article-title":"Functional analysis of conserved gene products involved in assembly of Escherichia coli capsules and exopolysaccharides: evidence for molecular recognition between Wza and Wzc for colanic acid biosynthesis","volume":"187","author":"Reid","year":"2005","journal-title":"J. Bacteriol."},{"key":"ref85","doi-asserted-by":"publisher","first-page":"515","DOI":"10.1007\/s00253-003-1517-y","article-title":"Gene expression in Escherichia coli biofilms","volume":"64","author":"Ren","year":"2004","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref86","doi-asserted-by":"publisher","first-page":"3048","DOI":"10.1128\/aac.00113-06","article-title":"Transcriptomic and functional analysis of an autolysis-deficient, teicoplanin-resistant derivative of methicillin-resistant Staphylococcus aureus","volume":"50","author":"Renzoni","year":"2006","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref87","doi-asserted-by":"publisher","first-page":"417","DOI":"10.1038\/s42003-019-0667-0","article-title":"Thermosensitive PBP2a requires extracellular folding factors PrsA and HtrA1 for Staphylococcus aureus MRSA \u03b2-lactam resistance","volume":"2","author":"Roch","year":"2019","journal-title":"Commun. Biol."},{"key":"ref88","doi-asserted-by":"publisher","first-page":"1754","DOI":"10.1128\/jb.182.6.1754-1756.2000","article-title":"AcrD of Escherichia coli is an aminoglycoside efflux pump","volume":"182","author":"Rosenberg","year":"2000","journal-title":"J. Bacteriol."},{"key":"ref89","doi-asserted-by":"publisher","first-page":"D255","DOI":"10.1093\/nar\/gkad1072","article-title":"RegulonDB v12.0: a comprehensive resource of transcriptional regulation in E. coli K-12","volume":"52","author":"Salgado","year":"2024","journal-title":"Nucleic Acids Res."},{"key":"ref90","doi-asserted-by":"publisher","first-page":"77266","DOI":"10.7554\/eLife.77266","article-title":"Flagellar energy costs across the tree of life","volume":"11","author":"Schavemaker","year":"2022","journal-title":"eLife"},{"key":"ref91","doi-asserted-by":"publisher","first-page":"839","DOI":"10.1261\/rna.053959.115","article-title":"How many biological replicates are needed in an RNA-seq experiment and which differential expression tool should you use?","volume":"22","author":"Schurch","year":"2016","journal-title":"RNA"},{"key":"ref92","doi-asserted-by":"publisher","first-page":"eadg3683","DOI":"10.1126\/sciadv.adg3683","article-title":"Peptidomimetic antibiotics disrupt the lipopolysaccharide transport bridge of drug-resistant Enterobacteriaceae","volume":"9","author":"Schuster","year":"2023","journal-title":"Sci. Adv."},{"key":"ref93","volume-title":"Experiments with Gene Fusions","author":"Silhavy","year":"1984"},{"key":"ref94","doi-asserted-by":"publisher","first-page":"a000414","DOI":"10.1101\/cshperspect.a000414","article-title":"The bacterial cell envelope","volume":"2","author":"Silhavy","year":"2010","journal-title":"Cold Spring Harb. Perspect. Biol."},{"key":"ref95","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1016\/0378-1119(87)90095-3","article-title":"Improved single and multicopy lac-based cloning vectors for protein and operon fusions","volume":"53","author":"Simons","year":"1987","journal-title":"Gene"},{"key":"ref96","doi-asserted-by":"publisher","first-page":"183839","DOI":"10.1016\/j.bbamem.2021.183839","article-title":"Linking dual mode of action of host defense antimicrobial peptide thanatin: structures, lipopolysaccharide and LptA(m) binding of designed analogs","volume":"1864","author":"Sinha","year":"2022","journal-title":"Biochim. Biophys. Acta Biomembr."},{"key":"ref97","doi-asserted-by":"publisher","first-page":"17795","DOI":"10.1038\/s41598-017-18102-6","article-title":"Structure and interactions of a host defense antimicrobial peptide thanatin in lipopolysaccharide micelles reveal mechanism of bacterial cell agglutination","volume":"7","author":"Sinha","year":"2017","journal-title":"Sci. Rep."},{"key":"ref98","doi-asserted-by":"publisher","first-page":"e00418-17","DOI":"10.1128\/JB.00418-17","article-title":"Distinctive roles for periplasmic proteases in the maintenance of essential outer membrane protein assembly","volume":"199","author":"Soltes","year":"2017","journal-title":"J. Bacteriol."},{"key":"ref99","doi-asserted-by":"publisher","first-page":"4460","DOI":"10.1128\/JB.00270-08","article-title":"Functional analysis of the protein machinery required for transport of lipopolysaccharide to the outer membrane of Escherichia coli","volume":"190","author":"Sperandeo","year":"2008","journal-title":"J. Bacteriol."},{"key":"ref100","doi-asserted-by":"publisher","first-page":"366","DOI":"10.1016\/j.resmic.2019.07.005","article-title":"The Lpt ABC transporter for lipopolysaccharide export to the cell surface","volume":"170","author":"Sperandeo","year":"2019","journal-title":"Res. Microbiol."},{"key":"ref101","doi-asserted-by":"publisher","first-page":"4885","DOI":"10.1128\/jb.178.16.4885-4893.1996","article-title":"Organization of the Escherichia coli K-12 gene cluster responsible for production of the extracellular polysaccharide colanic acid","volume":"178","author":"Stevenson","year":"1996","journal-title":"J. Bacteriol."},{"key":"ref102","doi-asserted-by":"publisher","first-page":"1403","DOI":"10.1111\/j.1365-2958.2005.04497.x","article-title":"Changes in lipopolysaccharide structure induce the sigma(E)-dependent response of Escherichia coli","volume":"55","author":"Tam","year":"2005","journal-title":"Mol. Microbiol."},{"key":"ref103","doi-asserted-by":"publisher","first-page":"16504","DOI":"10.1073\/pnas.1212797109","article-title":"Discovery of a cardiolipin synthase utilizing phosphatidylethanolamine and phosphatidylglycerol as substrates","volume":"109","author":"Tan","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref104","doi-asserted-by":"publisher","first-page":"a025288","DOI":"10.1101\/cshperspect.a025288","article-title":"Polymyxin: alternative mechanisms of action and resistance","volume":"6","author":"Trimble","year":"2016","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"ref105","doi-asserted-by":"publisher","first-page":"9","DOI":"10.1016\/j.molcel.2015.05.011","article-title":"Obg and membrane depolarization are part of a microbial bet-hedging strategy that leads to antibiotic tolerance","volume":"59","author":"Verstraeten","year":"2015","journal-title":"Mol. Cell"},{"key":"ref106","doi-asserted-by":"publisher","first-page":"eaau2634","DOI":"10.1126\/sciadv.aau2634","article-title":"Thanatin targets the intermembrane protein complex required for lipopolysaccharide transport in Escherichia coli","volume":"4","author":"Vetterli","year":"2018","journal-title":"Sci. Adv."},{"key":"ref107","doi-asserted-by":"publisher","first-page":"311","DOI":"10.1006\/jmbi.2000.4217","article-title":"Relationship between exopolysaccharide production and protein-tyrosine phosphorylation in gram-negative bacteria","volume":"304","author":"Vincent","year":"2000","journal-title":"J. Mol. Biol."},{"key":"ref108","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1146\/annurev-micro-090817-062640","article-title":"The complex Rcs regulatory Cascade","volume":"72","author":"Wall","year":"2018","journal-title":"Ann. Rev. Microbiol."},{"key":"ref109","doi-asserted-by":"publisher","first-page":"570","DOI":"10.1099\/mic.0.033415-0","article-title":"Different effects of transcriptional regulators MarA, SoxS and rob on susceptibility of Escherichia coli to cationic antimicrobial peptides (CAMPs): rob-dependent CAMP induction of the marRAB operon","volume":"156","author":"Warner","year":"2010","journal-title":"Microbiology"},{"key":"ref110","doi-asserted-by":"publisher","first-page":"7013","DOI":"10.1074\/jbc.275.10.7013","article-title":"The RcsAB box. Characterization of a new operator essential for the regulation of exopolysaccharide biosynthesis in enteric bacteria","volume":"275","author":"Wehland","year":"2000","journal-title":"J. Biol. Chem."},{"key":"ref111","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1038\/nprot.2007.521","article-title":"Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances","volume":"3","author":"Wiegand","year":"2008","journal-title":"Nat. Protoc."},{"key":"ref112","doi-asserted-by":"publisher","first-page":"e00836-21","DOI":"10.1128\/mBio.00836-21","article-title":"DpaA detaches Braun's lipoprotein from peptidoglycan","volume":"12","author":"Winkle","year":"2021","journal-title":"mBio"},{"key":"ref113","doi-asserted-by":"publisher","first-page":"744","DOI":"10.1016\/0300-9084(96)88192-2","article-title":"A family of homologous substrate-binding proteins with a broad range of substrate specificity and dissimilar biological functions","volume":"77","author":"Wu","year":"1995","journal-title":"Biochimie"},{"key":"ref114","doi-asserted-by":"publisher","first-page":"6368","DOI":"10.1038\/s41467-023-42007-w","article-title":"LptM promotes oxidative maturation of the lipopolysaccharide translocon by substrate binding mimicry","volume":"14","author":"Yang","year":"2023","journal-title":"Nat. Commun."},{"key":"ref115","doi-asserted-by":"publisher","first-page":"389","DOI":"10.1038\/415389a","article-title":"Antimicrobial peptides of multicellular organisms","volume":"415","author":"Zasloff","year":"2002","journal-title":"Nature"},{"key":"ref116","doi-asserted-by":"publisher","first-page":"18","DOI":"10.1186\/1471-2199-12-18","article-title":"Novel reference genes for quantifying transcriptional responses of Escherichia coli to protein overexpression by quantitative PCR","volume":"12","author":"Zhou","year":"2011","journal-title":"BMC Mol. Biol."}],"container-title":["Frontiers in Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2026.1920608\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T06:00:48Z","timestamp":1790748048000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2026.1920608\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,30]]},"references-count":116,"alternative-id":["10.3389\/fmicb.2026.1920608"],"URL":"https:\/\/doi.org\/10.3389\/fmicb.2026.1920608","relation":{},"ISSN":["1664-302X"],"issn-type":[{"value":"1664-302X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,30]]},"article-number":"1920608"}}