{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T13:55:17Z","timestamp":1790776517107,"version":"4.1.0"},"reference-count":43,"publisher":"American Society for Microbiology","license":[{"start":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T00:00:00Z","timestamp":1790726400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T00:00:00Z","timestamp":1790726400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      IncI2-type plasmids are major vehicles for the dissemination of\n                      <jats:italic toggle=\"yes\">mcr-1<\/jats:italic>\n                      . These plasmids encode a type IV secretion system (T4SS) that mediates conjugative transfer and a type IV pilus (T4P) that facilitates mating-pair formation. Shufflon-mediated diversity of tip adhesin PilV of T4P may contribute to the extensive dissemination of IncI2 plasmids. Herein, we identified a novel shufflon segment forming an active new PilV variant from an IncI2 plasmid, which was named PilV-E and was detected in 127 of 501 (25.35%) complete IncI2 plasmids in public databases. We further demonstrated that strains harboring the single PilV-E variant displayed a distinct functional profile, characterized by both robust biofilm formation and high\n                      <jats:italic toggle=\"yes\">Caco-2<\/jats:italic>\n                      adhesion. Compared with the \u0394\n                      <jats:italic toggle=\"yes\">pilV<\/jats:italic>\n                      -shufflon-\n                      <jats:italic toggle=\"yes\">rci<\/jats:italic>\n                      strain, the PilV-E-carrying strain showed increased biofilm formation (OD\n                      <jats:sub>590<\/jats:sub>\n                      from 1.50 to 2.60), increased\n                      <jats:italic toggle=\"yes\">Caco-2<\/jats:italic>\n                      adhesion (0.04% to 5.46%), and increased surface motility (1.20 to 1.87 cm\/day). Among all variants, PilV-A' showed the strongest cell adhesion (6.00%), whereas PilV-B' showed the greatest motility (2.54 cm\/day). Notably, PilV-E significantly restored conjugation frequency compared with the \u0394\n                      <jats:italic toggle=\"yes\">pilV<\/jats:italic>\n                      -shufflon-\n                      <jats:italic toggle=\"yes\">rci<\/jats:italic>\n                      strain and displayed the highest and lowest frequencies when recipients were\n                      <jats:italic toggle=\"yes\">Escherichia coli<\/jats:italic>\n                      (5.23 \u00d7 10\n                      <jats:sup>\u22123<\/jats:sup>\n                      ) and\n                      <jats:italic toggle=\"yes\">Acinetobacter baumannii<\/jats:italic>\n                      (4.46 \u00d7 10\n                      <jats:sup>\u22128<\/jats:sup>\n                      ), respectively. All findings confirm that PilV-E serves as a novel and key tip adhesin of T4P and plays a crucial role in plasmid conjugation, indicating that the diversity of shufflon-mediated PilV variants has been underestimated and providing insight into the molecular basis for the extensive spread of IncI2 plasmids.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        The plasmid-mediated colistin resistance gene\n                        <jats:italic toggle=\"yes\">mcr-1<\/jats:italic>\n                        has become a major public health concern because it compromises one of the last-line treatments for multidrug-resistant Gram-negative bacterial infections. IncI2 plasmids are among the most important vehicles for the dissemination of\n                        <jats:italic toggle=\"yes\">mcr-1<\/jats:italic>\n                        , but the molecular factors that facilitate their efficient transfer remain insufficiently understood. In this study, we identified PilV-E, a novel shufflon-associated type IV pilus tip adhesin encoded by a human-derived IncI2 plasmid, and demonstrated its contribution to bacterial fitness-related phenotypes and conjugative transfer. The prevalence of PilV-E among complete IncI2 plasmids further indicates that this variant has been overlooked in previous studies. These findings expand our understanding of shufflon-mediated PilV diversity and provide new insight into how IncI2 plasmids participate in the efficient dissemination of\n                        <jats:italic toggle=\"yes\">mcr-1<\/jats:italic>\n                        among diverse bacterial species, including clinically important pathogens.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.01825-26","type":"journal-article","created":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T13:00:21Z","timestamp":1790773221000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Discovery and functional characterization of a novel PilV variant in a\n                    <i>mcr-1<\/i>\n                    -positive IncI2 plasmid"],"prefix":"10.1128","author":[{"given":"Chen","family":"An","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04v3ywz14","id-type":"ROR","asserted-by":"publisher"}],"name":"State Key Laboratory of Veterinary Public Health and Safety, College of Veterinary Medicine, China Agricultural University","place":["Beijing, China"]},{"id":[{"id":"https:\/\/ror.org\/04v3ywz14","id-type":"ROR","asserted-by":"publisher"}],"name":"College of Veterinary Medicine, China Agricultural University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Yang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04v3ywz14","id-type":"ROR","asserted-by":"publisher"}],"name":"College of Veterinary Medicine, China Agricultural University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silin","family":"Liu","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04v3ywz14","id-type":"ROR","asserted-by":"publisher"}],"name":"Sanya Institute, China Agricultural University","place":["Sanya, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dongyan","family":"Shao","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04v3ywz14","id-type":"ROR","asserted-by":"publisher"}],"name":"State Key Laboratory of Veterinary Public Health and Safety, College of Veterinary Medicine, China Agricultural University","place":["Beijing, China"]},{"id":[{"id":"https:\/\/ror.org\/04v3ywz14","id-type":"ROR","asserted-by":"publisher"}],"name":"College of Veterinary Medicine, China Agricultural University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9478-4023","authenticated-orcid":true,"given":"Hongwei","family":"Zhou","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00a2xv884","id-type":"ROR","asserted-by":"publisher"}],"name":"The Second Affiliated Hospital Zhejiang University School of Medicine","place":["Hangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuchen","family":"Wu","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00a2xv884","id-type":"ROR","asserted-by":"publisher"}],"name":"The Second Affiliated Hospital Zhejiang University School of Medicine","place":["Hangzhou, 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standard M07","author":"Clinical and Laboratory Standards Institute","year":"2024","unstructured":"Clinical and Laboratory Standards Institute. 2024. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically. In CLSI standard M07, 12th ed. Clinical and Laboratory Standards Institute, Berwyn, PA, USA.","edition":"12"},{"key":"e_1_3_4_43_2","volume-title":"CLSI supplement M100","author":"Clinical and Laboratory Standards Institute","year":"2026","unstructured":"Clinical and Laboratory Standards Institute. 2026. Performance standards for antimicrobial susceptibility testing. In CLSI supplement M100, 36th ed. Clinical and Laboratory Standards Institute, Berwyn, PA, USA.","edition":"36"},{"key":"e_1_3_4_44_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-024-07487-w"}],"container-title":["Microbiology Spectrum"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.01825-26","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.01825-26","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T13:00:22Z","timestamp":1790773222000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/spectrum.01825-26"}},"subtitle":[],"editor":[{"given":"Jennifer M.","family":"Auchtung","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,9,30]]},"references-count":43,"alternative-id":["10.1128\/spectrum.01825-26"],"URL":"https:\/\/doi.org\/10.1128\/spectrum.01825-26","relation":{},"ISSN":["2165-0497"],"issn-type":[{"value":"2165-0497","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,30]]},"assertion":[{"value":"2026-05-29","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-08","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-30","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e01825-26"}}