{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T14:33:48Z","timestamp":1787236428241,"version":"build-2736575974"},"reference-count":48,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T00:00:00Z","timestamp":1787184000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Key R&D Program of China during the 14th Five year Plan Period","award":["2022YFD1302101"],"award-info":[{"award-number":["2022YFD1302101"]}]},{"name":"Key Research and Development Program of Gansu Province","award":["26YFNA007"],"award-info":[{"award-number":["26YFNA007"]}]},{"name":"Project supported by the Natural Science Foundation of Gansu Province","award":["24JRRA022"],"award-info":[{"award-number":["24JRRA022"]}]},{"name":"Special Project of the Natural Science Foundation of Gansu Province, China","award":["26JRRA380"],"award-info":[{"award-number":["26JRRA380"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vet. Sci.","Veterinary Sciences"],"abstract":"<jats:p>(1) Background: methicillin\u2011resistant Staphylococcus aureus (MRSA) is a zoonotic pathogen, and its hemolysins serve as key virulence factors. Functional membrane microdomains (FMMs) are protein-enriching platforms and regulate diverse physiological functions by recruiting and assembling various proteins. However, whether and how FMMs regulate the hemolytic ability of MRSA remains unclear. This study aimed to investigate FMM-mediated regulation of MRSA hemolysins and the underlying mechanisms. (2) Methods: Homologous recombination was employed to generate FMM-disrupted (N315\u0394floA) and complemented (N315\u0394floA::floA) strains from the MRSA N315 wild-type strain (N315 WT). The three strains were compared with respect to hemolytic activity, transcript levels of key virulence and regulatory genes, and in vivo virulence. (3) Results: Disruption of FMMs significantly enhanced hemolytic activity compared with N315 WT and complemented strains. Meanwhile, FMM disruption repressed the two-component system genes (vraS and vraR), while activating the agr operon (agrB, agrD, agrC and agrA) and its effector molecule RNAIII, leading to upregulation of hemolysin genes (hla, hlb, hld). In vivo, N315\u0394floA infection markedly increased mortality in G. mellonella larvae and BALB\/c mice, with significantly elevated pro-inflammatory factors (TNF-\u03b1, IL-6, and IL-1\u03b2) in mouse plasma. All these phenotypes were effectively reversed in N315\u0394floA::floA. (4) Conclusions: Disruption of FMMs potentiates both hemolytic activity and overall virulence in MRSA, with the potential underlying mechanism involving the VraS\/R-Agr regulatory axis that drives transcriptional upregulation of hemolysin-encoding genes.<\/jats:p>","DOI":"10.3390\/vetsci13080839","type":"journal-article","created":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T12:57:53Z","timestamp":1787230673000},"page":"839","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Disruption of Functional Membrane Microdomains Enhances Methicillin-Resistant Staphylococcus aureus Pathogenesis via Hyperexpression of Hemolysins"],"prefix":"10.3390","volume":"13","author":[{"given":"Bingtian","family":"Jin","sequence":"first","affiliation":[{"name":"Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China"},{"name":"The First School of Clinical Medicine, Gansu University of Chinese Medicine, Lanzhou 730000, China"},{"name":"Key Laboratory of Veterinary Drug Discovery, Ministry of Agriculture and Rural Affairs, Lanzhou 730050, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-4944-6609","authenticated-orcid":false,"given":"Changzhen","family":"Wang","sequence":"additional","affiliation":[{"name":"Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China"},{"name":"Key Laboratory of Veterinary Drug Discovery, Ministry of Agriculture and Rural Affairs, Lanzhou 730050, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tiantian","family":"Liu","sequence":"additional","affiliation":[{"name":"Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China"},{"name":"Key Laboratory of Veterinary Drug Discovery, Ministry of Agriculture and Rural Affairs, Lanzhou 730050, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pengcheng","family":"Dong","sequence":"additional","affiliation":[{"name":"Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China"},{"name":"The First School of Clinical Medicine, Gansu University of Chinese Medicine, Lanzhou 730000, China"},{"name":"Key Laboratory of Veterinary Drug Discovery, Ministry of Agriculture and Rural Affairs, Lanzhou 730050, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xurong","family":"Wang","sequence":"additional","affiliation":[{"name":"Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China"},{"name":"Key Laboratory of Veterinary Drug Discovery, Ministry of Agriculture and Rural Affairs, Lanzhou 730050, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiao","family":"Yang","sequence":"additional","affiliation":[{"name":"Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China"},{"name":"Key Laboratory of Veterinary Drug Discovery, Ministry of Agriculture and Rural Affairs, Lanzhou 730050, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dengwang","family":"Yuan","sequence":"additional","affiliation":[{"name":"Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China"},{"name":"Key Laboratory of Veterinary Drug Discovery, Ministry of Agriculture and Rural Affairs, Lanzhou 730050, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Feng","family":"Yang","sequence":"additional","affiliation":[{"name":"Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China"},{"name":"Key Laboratory of Veterinary Drug Discovery, Ministry of Agriculture and Rural Affairs, Lanzhou 730050, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"11251","DOI":"10.1038\/s41467-025-66118-8","article-title":"Vancomycin heteroresistance (hVISA) in MRSA links to treatment failure and supports a revised PAP-AUC threshold","volume":"16","author":"Kim","year":"2025","journal-title":"Nat. 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