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However, the functional packing and precise role of VirB8 in T4SSs biology remain undefined. Our findings elucidate the nature of VirB8 proteins as cell wall components, where they multimerize and exhibit a conserved assembly pattern, distinct from VirB8 in Gram-negative bacteria. We also demonstrate that VirB8 proteins interact with other T4SS subunits and DNA, indicating their pivotal role in the building of the DNA translocation channel across the cell wall. We lastly propose a distinct architecture for conjugative T4SSs in Gram-positive bacteria compared to their Gram-negative counterparts, possibly attributed to the differences in the cell wall structure.<\/jats:p>","DOI":"10.1126\/sciadv.adq5975","type":"journal-article","created":{"date-parts":[[2025,1,22]],"date-time":"2025-01-22T18:58:39Z","timestamp":1737572319000},"update-policy":"https:\/\/doi.org\/10.34133\/aaas_crossmark","source":"Crossref","is-referenced-by-count":7,"title":["Elucidating assembly and function of VirB8 cell wall subunits refines the DNA translocation model in Gram-positive T4SSs"],"prefix":"10.1126","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2746-0229","authenticated-orcid":true,"given":"Robine","family":"Maffo-Woulefack","sequence":"first","affiliation":[{"name":"Universit\u00e9 de Lorraine, INRAE, DynAMic, F-54000 Nancy, France."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abbas 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