{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T02:34:54Z","timestamp":1792118094743,"version":"4.4.0"},"update-to":[{"DOI":"10.1371\/journal.ppat.1013474","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2025,9,11]],"date-time":"2025-09-11T00:00:00Z","timestamp":1757548800000}}],"reference-count":95,"publisher":"Public Library of Science (PLoS)","issue":"9","license":[{"start":{"date-parts":[[2025,9,5]],"date-time":"2025-09-05T00:00:00Z","timestamp":1757030400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005416","name":"Norges Forskningsr\u00c3\u00a5d","doi-asserted-by":"publisher","award":["223255\/F50"],"award-info":[{"award-number":["223255\/F50"]}],"id":[{"id":"10.13039\/501100005416","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005416","name":"Norges Forskningsr\u00c3\u00a5d","doi-asserted-by":"publisher","award":["249901"],"award-info":[{"award-number":["249901"]}],"id":[{"id":"10.13039\/501100005416","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001704","name":"European Society of Clinical Microbiology and Infectious Diseases","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001704","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010434","name":"'la Caixa' Foundation","doi-asserted-by":"publisher","award":["LCF\/PR\/GN16\/10290002"],"award-info":[{"award-number":["LCF\/PR\/GN16\/10290002"]}],"id":[{"id":"10.13039\/100010434","id-type":"DOI","asserted-by":"publisher"}]},{"name":"CIBER Enfermedades Respiratorias"},{"name":"Spanish Government-FEDER Funds","award":["PI17\/01511"],"award-info":[{"award-number":["PI17\/01511"]}]}],"content-domain":{"domain":["www.plospathogens.org"],"crossmark-restriction":false},"short-container-title":["PLoS Pathog"],"abstract":"<jats:p><jats:italic><jats:italic>Drosophila melanogaster<\/jats:italic><\/jats:italic> (<jats:italic><jats:italic>Drosophila<\/jats:italic><\/jats:italic>) is one of the most extensively studied animal models we have, with a broad, advanced, and organized research community. Yet, <jats:italic><jats:italic>Drosophila<\/jats:italic><\/jats:italic> has barely been exploited to understand the underlying mechanisms of mycobacterial infections, which cause some of the deadliest infectious diseases humans are currently battling. Here, we identified mycobacterial genes required for the pathogen\u2019s growth during <jats:italic><jats:italic>Drosophila<\/jats:italic><\/jats:italic> infection. Using <jats:italic><jats:italic>Mycobacterium marinum<\/jats:italic><\/jats:italic> (<jats:italic><jats:italic>Mmar<\/jats:italic><\/jats:italic>) to model mycobacterial pathogens, we first validated that an established mycobacterial virulence factor, EccB1 of the ESX-1 Type VII secretion system, is required for <jats:italic><jats:italic>Mmar<\/jats:italic><\/jats:italic> growth within the flies. Subsequently, we identified <jats:italic><jats:italic>Mmar<\/jats:italic><\/jats:italic> virulence genes in <jats:italic><jats:italic>Drosophila<\/jats:italic><\/jats:italic> in a high-throughput genome-wide phenotypic manner using transposon insertion sequencing. Of the 181 identified virulence genes, the vast majority (91%) had orthologs in the tuberculosis-causing <jats:italic>M. tuberculosis<\/jats:italic> (<jats:italic>Mtb<\/jats:italic>), suggesting that the encoded virulence mechanisms may be conserved across <jats:italic><jats:italic>Mmar<\/jats:italic><\/jats:italic> and <jats:italic><jats:italic>Mtb<\/jats:italic><\/jats:italic> species. By studying one of the identified genes in more depth, the putative ATP-binding protein ABC transporter encoded by <jats:italic><jats:italic>mmar_1660<\/jats:italic><\/jats:italic>, we found that both the <jats:italic><jats:italic>Mmar<\/jats:italic><\/jats:italic> gene and its <jats:italic><jats:italic>Mtb<\/jats:italic><\/jats:italic> ortholog (<jats:italic><jats:italic>rv3041c<\/jats:italic><\/jats:italic>) were required for virulence in human macrophages as well. We pinpointed the probable virulence mechanism of the genes to their requirements for growth during iron limitation, a condition met by mycobacteria during host infection. Together, our results bring forward <jats:italic>Drosophila<\/jats:italic> as a promising host model to study and identify mycobacterial virulence factors, providing insights that may transfer to <jats:italic><jats:italic>Mtb<\/jats:italic><\/jats:italic> human infection.<\/jats:p>","DOI":"10.1371\/journal.ppat.1013474","type":"journal-article","created":{"date-parts":[[2025,9,5]],"date-time":"2025-09-05T17:51:32Z","timestamp":1757094692000},"page":"e1013474","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":4,"title":["Genome-wide phenotypic insights into mycobacterial virulence using Drosophila melanogaster"],"prefix":"10.1371","volume":"21","author":[{"given":"Niruja","family":"Sivakumar","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Esther","family":"Fuentes","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Linn-Karina","family":"Selvik","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marta","family":"Arch","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christina Gabrielsen","family":"\u00c5s","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pere-Joan","family":"Cardona","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas R.","family":"Ioerger","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1546-4329","authenticated-orcid":true,"given":"Marte Sings\u00e5s","family":"Dragset","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2025,9,5]]},"reference":[{"key":"ppat.1013474.ref001","author":"WHO"},{"issue":"11","key":"ppat.1013474.ref002","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1038\/s41579-022-00731-y","article-title":"Anti-tuberculosis treatment strategies and drug development: challenges and priorities","volume":"20","author":"VA Dartois","year":"2022","journal-title":"Nat Rev Microbiol"},{"issue":"1","key":"ppat.1013474.ref003","doi-asserted-by":"crossref","first-page":"418","DOI":"10.1186\/1471-2458-14-418","article-title":"Tuberculosis treatment discontinuation and symptom persistence: an observational 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