{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:18:44Z","timestamp":1785403124068,"version":"3.56.0"},"reference-count":32,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2026,4,5]],"date-time":"2026-04-05T00:00:00Z","timestamp":1775347200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32070188"],"award-info":[{"award-number":["32070188"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"award":["32070188"],"award-info":[{"award-number":["32070188"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Mycoplasma pneumoniae is a significant pathogen responsible for community-acquired respiratory infections in children and adolescents, with the rising prevalence of macrolide-resistant M. pneumoniae (MRMP), particularly in Asia, presenting critical treatment challenges. Our previous study inferred that a macrolide efflux pump may contribute to macrolide resistance in M. pneumoniae in addition to the common point mutations in 23S rRNA gene. This study aimed to define the specific pump and confirm its role. Through comparative genomic analysis, we identified a candidate gene, MPN_080, encoding an ABC transporter permease, which was further characterized using phylogenetic analysis, AlphaFold-based structural modeling, and biochemical assays. Overexpression of MPN_080 from an erythromycin-resistant isolate in the erythromycin-sensitive M129 resulted in a significant increase in minimum inhibitory concentrations (MICs) from &lt;0.125 \u00b5g\/mL to 1 \u00b5g\/mL, while similar overexpression of MPN_080 derived from M129 did not affect MICs. Notably, this resistance mechanism operates independently of M. pneumoniae virulence factors, as evidenced by unaltered colonization capacity in NCI-H292 cells and consistent immune response patterns across both strains. Our findings establish MPN_080 as a novel determinant of macrolide resistance functioning associated with enhanced ATPase activity. These insights into non-classical resistance mechanisms may guide future diagnostic and therapeutic strategies against MRMP.<\/jats:p>","DOI":"10.3390\/microorganisms14040831","type":"journal-article","created":{"date-parts":[[2026,4,6]],"date-time":"2026-04-06T04:10:27Z","timestamp":1775448627000},"page":"831","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Beyond Ribosomal Mutations: Identification of MPN_080 as a Novel ATPase-Dependent Determinant of Macrolide Resistance in Mycoplasma pneumoniae"],"prefix":"10.3390","volume":"14","author":[{"given":"Shaoli","family":"Li","sequence":"first","affiliation":[{"name":"Capital Institute of Pediatrics-Peking University Teaching Hospital, Beijing 100020, China"},{"name":"Capital Center for Children\u2019s Health, Capital Medical University, Beijing 100020, China"},{"name":"Capital Center for Children\u2019s Health, Capital Institute of Pediatrics, Capital Medical University, Beijing 100020, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-2365-0641","authenticated-orcid":false,"given":"Yuyan","family":"Xia","sequence":"additional","affiliation":[{"name":"Capital Center for Children\u2019s Health, Capital Institute of Pediatrics, Capital Medical University, Beijing 100020, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fei","family":"Zhao","sequence":"additional","affiliation":[{"name":"National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5143-0826","authenticated-orcid":false,"given":"Xiuwei","family":"Wang","sequence":"additional","affiliation":[{"name":"Capital Center for Children\u2019s Health, Capital Institute of Pediatrics, Capital Medical University, Beijing 100020, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhengli","family":"Li","sequence":"additional","affiliation":[{"name":"Capital Center for Children\u2019s Health, Capital Institute of Pediatrics, Capital Medical University, Beijing 100020, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liyong","family":"Liu","sequence":"additional","affiliation":[{"name":"National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junting","family":"Liu","sequence":"additional","affiliation":[{"name":"Capital Center for Children\u2019s Health, Capital Institute of Pediatrics, Capital Medical University, Beijing 100020, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mei","family":"Diao","sequence":"additional","affiliation":[{"name":"Capital Institute of Pediatrics-Peking University Teaching Hospital, Beijing 100020, China"},{"name":"Capital Center for Children\u2019s Health, Capital Medical University, Beijing 100020, China"},{"name":"Capital Center for Children\u2019s Health, Capital Institute of Pediatrics, Capital Medical University, Beijing 100020, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"6343818","DOI":"10.1155\/2022\/6343818","article-title":"Mycoplasma pneumoniae Infections and Primary Immune Deficiencies","volume":"2022","author":"Poddighe","year":"2022","journal-title":"Int. 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