{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T20:27:02Z","timestamp":1792787222895,"version":"4.5.0"},"reference-count":68,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2024,8,7]],"date-time":"2024-08-07T00:00:00Z","timestamp":1722988800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2024,8,7]],"date-time":"2024-08-07T00:00:00Z","timestamp":1722988800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"name":"EC | Horizon Europe | Excellent Science | HORIZON EUROPE Marie Sklodowska-Curie Actions","award":["956154"],"award-info":[{"award-number":["956154"]}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2024,8,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      Intrinsic resistance to macrolides in Gram-negative bacteria is primarily attributed to the low permeability of the outer membrane, though the underlying genetic and molecular mechanisms remain to be fully elucidated. Here, we used transposon directed insertion-site sequencing (TraDIS) to identify chromosomal non-essential genes involved in\n                      <jats:italic>Escherichia coli<\/jats:italic>\n                      intrinsic resistance to a macrolide antibiotic, tilmicosin. We constructed two highly saturated transposon mutant libraries of &gt;290,000 and &gt;390,000 unique Tn5 insertions in a clinical enterotoxigenic strain (ETEC5621) and in a laboratory strain (K-12 MG1655), respectively. TraDIS analysis identified genes required for growth of ETEC5621 and MG1655 under 1\/8 MIC (\n                      <jats:italic>n<\/jats:italic>\n                      = 15 and 16, respectively) and 1\/4 MIC (\n                      <jats:italic>n<\/jats:italic>\n                      = 38 and 32, respectively) of tilmicosin. For both strains, 23 genes related to lipopolysaccharide biosynthesis, outer membrane assembly, the Tol-Pal system, efflux pump, and peptidoglycan metabolism were enriched in the presence of the antibiotic. Individual deletion of genes (\n                      <jats:italic>n<\/jats:italic>\n                      = 10) in the wild-type strains led to a 64- to 2-fold reduction in MICs of tilmicosin, erythromycin, and azithromycin, validating the results of the TraDIS analysis. Notably, deletion of\n                      <jats:italic>surA<\/jats:italic>\n                      or\n                      <jats:italic>waaG<\/jats:italic>\n                      , which impairs the outer membrane, led to the most significant decreases in MICs of all three macrolides in ETEC5621. Our findings contribute to a genome-wide understanding of intrinsic macrolide resistance in\n                      <jats:italic>E. coli<\/jats:italic>\n                      , shedding new light on the potential role of the peptidoglycan layer. They also provide an\n                      <jats:italic>in vitro<\/jats:italic>\n                      proof of concept that\n                      <jats:italic>E. coli<\/jats:italic>\n                      can be sensitized to macrolides by targeting proteins maintaining the outer membrane such as SurA and WaaG.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/aac.00452-24","type":"journal-article","created":{"date-parts":[[2024,6,28]],"date-time":"2024-06-28T09:01:39Z","timestamp":1719565299000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":24,"title":["The intrinsic macrolide resistome of\n                    <i>Escherichia coli<\/i>"],"prefix":"10.1128","volume":"68","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-0143-7477","authenticated-orcid":false,"given":"Yibing","family":"Ma","sequence":"first","affiliation":[{"name":"Department of Veterinary and Animal Sciences, University of Copenhagen, Frederiksberg, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9814-1500","authenticated-orcid":false,"given":"Mattia","family":"Pirolo","sequence":"additional","affiliation":[{"name":"Department of Veterinary and Animal Sciences, University of Copenhagen, Frederiksberg, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bimal","family":"Jana","sequence":"additional","affiliation":[{"name":"Center for Genomic Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Viktor Hundtofte","family":"Mebus","sequence":"additional","affiliation":[{"name":"Department of Veterinary and Animal Sciences, University of Copenhagen, Frederiksberg, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2389-5084","authenticated-orcid":true,"given":"Luca","family":"Guardabassi","sequence":"additional","affiliation":[{"name":"Department of Veterinary and Animal Sciences, University of Copenhagen, Frederiksberg, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1111\/bph.13936"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.37.2.354"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.3109\/1040841X.2015.1136261"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/msphere.00402-22"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-019-42423-3"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid1510.090696"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41576-020-0244-x"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1101\/gr.097097.109"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.02096-17"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics12060993"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1038\/srep42483"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.00287-22"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.174.8.2525-2538.1992"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00598-20"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1007\/s002030050732"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.184.2.363-369.2002"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.144709"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.178.6.1770-1773.1996"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro2069"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2011.07539.x"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1312012110"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.5483\/BMBRep.2023-0070"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1093\/femsre\/fuaa018"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2022.878049"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1507760112"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmcl.2018.09.034"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics5010010"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.184.23.6490-6499.2002"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1038\/ja.2016.50"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-020-0366-3"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2300784120"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1919267117"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-022-04834-7"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2100370118"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0120060"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.029553-0"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01114-17"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.1007\/s40262-014-0209-3"},{"key":"e_1_3_4_40_2","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.01205-23"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00117-14"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.176.13.4144-4156.1994"},{"key":"e_1_3_4_43_2","doi-asserted-by":"publisher","DOI":"10.1177\/104063870201400505"},{"key":"e_1_3_4_44_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2019.01.032"},{"key":"e_1_3_4_45_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bpj.2015.06.036"},{"key":"e_1_3_4_46_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2019.00100"},{"key":"e_1_3_4_47_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.gene.2023.147943"},{"key":"e_1_3_4_48_2","doi-asserted-by":"publisher","DOI":"10.1093\/jtm\/tax026"},{"key":"e_1_3_4_49_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.04729-14"},{"key":"e_1_3_4_50_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkx268"},{"key":"e_1_3_4_51_2","doi-asserted-by":"publisher","DOI":"10.1128\/ecosalplus.ESP-0006-2016"},{"key":"e_1_3_4_52_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.envint.2018.02.007"},{"key":"e_1_3_4_53_2","unstructured":"European medicines agency. 2019. Categorisation of antibiotics in the European Union. Available from: https:\/\/www.ema.europa.eu\/en\/documents\/report\/categorisation-antibiotics-european-union-answer-request-european-commission-updating-scientific-advice-impact-public-health-and-animal-health-use-antibiotics-animals_en.pdf. Retrieved 19 Mar 2024."},{"key":"e_1_3_4_54_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.prevetmed.2023.105852"},{"key":"e_1_3_4_55_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.00559-23"},{"key":"e_1_3_4_56_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.micpath.2020.104389"},{"key":"e_1_3_4_57_2","doi-asserted-by":"publisher","DOI":"10.1038\/msb4100050"},{"key":"e_1_3_4_58_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.01913-20"},{"key":"e_1_3_4_59_2","volume-title":"CLSI supplement M100Performance standards for antimicrobial susceptibility testing","author":"CLSI","year":"2018","unstructured":"CLSI. 2018. CLSI supplement M100. Performance standards for antimicrobial susceptibility testing. 28th ed. Clinical and Laboratory Standards Institute, Wayne, PA.","edition":"28"},{"key":"e_1_3_4_60_2","volume-title":"Methods for determining bactericidal activity of antimicrobial agents; approved guideline","author":"CLSI","year":"1999","unstructured":"CLSI. 1999. Methods for determining bactericidal activity of antimicrobial agents; approved guideline. Clinical and Laboratory Standards Institute, Wayne, PA."},{"key":"e_1_3_4_61_2","doi-asserted-by":"publisher","DOI":"10.1128\/9781555818814.ch5.16"},{"key":"e_1_3_4_62_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1005595"},{"key":"e_1_3_4_63_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btu153"},{"key":"e_1_3_4_64_2","doi-asserted-by":"publisher","DOI":"10.1101\/pdb.prot101220"},{"key":"e_1_3_4_65_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-022-15997-8"},{"key":"e_1_3_4_66_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btw022"},{"key":"e_1_3_4_67_2","doi-asserted-by":"publisher","DOI":"10.1099\/mgen.0.000719"},{"key":"e_1_3_4_68_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.120163297"},{"key":"e_1_3_4_69_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mimet.2008.06.010"}],"container-title":["Antimicrobial Agents and Chemotherapy"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/aac.00452-24","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/aac.00452-24","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,8,7]],"date-time":"2024-08-07T09:02:20Z","timestamp":1723021340000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/aac.00452-24"}},"subtitle":[],"editor":[{"given":"Boudewijn L.","family":"de Jonge","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2024,8,7]]},"references-count":68,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2024,8,7]]}},"alternative-id":["10.1128\/aac.00452-24"],"URL":"https:\/\/doi.org\/10.1128\/aac.00452-24","relation":{},"ISSN":["0066-4804","1098-6596"],"issn-type":[{"value":"0066-4804","type":"print"},{"value":"1098-6596","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,8,7]]},"assertion":[{"value":"2024-03-25","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-05-16","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-06-28","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e00452-24"}}