{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T01:06:06Z","timestamp":1789088766700,"version":"build-2803163510"},"reference-count":54,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T00:00:00Z","timestamp":1787011200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T00:00:00Z","timestamp":1787011200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32272986"],"award-info":[{"award-number":["32272986"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"2115 Talent Development Program of China Agricultural University"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Virol"],"published-print":{"date-parts":[[2026,8,18]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>Multibasic cleavage motifs play critical roles in envelope glycoprotein-mediated viral entry by serving as substrates for proprotein convertases like furin. The gamma-coronavirus infectious bronchitis virus (IBV) inherently possesses a conserved S1\/S2 multibasic cleavage site (MBCS) within its S gene. Notably, the IBV Beaudette strain uniquely harbors an additional S2\u2032 MBCS, which extends its cellular tropism. Our previous work demonstrated that introducing the S2\u2032 MBCS into the YN strain contributes to central nervous system damage. However, the functional interplay between the S1\/S2 and S2\u2032 MBCS has not yet been elucidated. Herein, we rescued several recombinant IBV mutants based on two different genotypes, engineered with distinct S protein cleavage patterns. Our results revealed that deletion of the S1\/S2 MBCS results in a drastic reduction of viral replication efficiency and pathogenicity. This negative effect can be partially compensated by introducing the S2\u2032 MBCS. The S2\u2032 MBCS also rendered increased mortality and elevated viral loads in neurological organs\u2014effects that were diminished with less efficient S1\/S2 cleavage. Inconsistencies exist regarding the impact of MBCS mutations in different IBV strain backgrounds, which are mainly due to their differential dependence on host proteases, correlating with their distinct tissue tropism. Our study demonstrated a proteolytic coordination model for IBV S protein activation: The S1\/S2 MBCS primarily confers efficient replication competence; the S2\u2032 site acts as an amplifier of organ-specific pathogenicity, particularly enhancing neurotropism. This synergistic relationship is conserved across different IBV strains.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      Multibasic cleavage sites (MBCSs) are critical determinants of coronavirus pathogenicity and tissue tropism. While infectious bronchitis virus (IBV) inherently possesses a conserved S1\/S2 cleavage site, the functional significance of this site and its potential interplay with the unique S2\u2032 MBCS found in the Beaudette strain remain poorly understood. Our study definitively establishes the critical role of the S1\/S2 cleavage site for efficient IBV replication\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      and full virulence\n                      <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                      , while the S2\u2032 site can only partially restore viral virulence. The pathogenic enhancement conferred by the S2\u2032 site is due to the heightened viral loads and enhanced organ-specific neurological damage. These findings provide essential insights into IBV pathogenesis governed by spike protein cleavage and establish a vital theoretical foundation for designing rationally attenuated vaccines and effective control strategies against avian coronaviruses.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/jvi.00614-26","type":"journal-article","created":{"date-parts":[[2026,7,6]],"date-time":"2026-07-06T13:22:28Z","timestamp":1783344148000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Disruption of the S1\/S2 multibasic cleavage site attenuates infectious bronchitis virus, while S2\u2032 partially restores viral virulence and expands tissue tropism"],"prefix":"10.1128","volume":"100","author":[{"given":"Ye","family":"Zhao","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":"Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun","family":"Zhao","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":"Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yingfei","family":"Li","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":"Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Linqing","family":"Duan","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":"Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rong","family":"Liang","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":"Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Min","family":"Huang","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":"Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-1985-4809","authenticated-orcid":true,"given":"Jing","family":"Zhao","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":"Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8200-312X","authenticated-orcid":true,"given":"Guozhong","family":"Zhang","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":"Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virusres.2006.01.017"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00705-003-0162-1"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1051\/vetres:2006055"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.27539"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2020.04.022"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.antiviral.2020.104742"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.21037\/jtd-21-226"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1080\/03079450500367682"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens12050698"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1017\/S1466252317000044"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1080\/03079457.2022.2063710"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2017.03.010"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01044-18"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2024.110134"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1407087111"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.3390\/v11100972"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02447-20"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00613-09"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.00535-25"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid1812.120552"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/0300-9084(94)90149-x"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.02327-24"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41580-021-00418-x"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1021\/acsinfecdis.0c00701"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1004502"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2021.649314"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41594-020-0468-7"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.02808-24"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1080\/22221751.2021.2014284"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1093\/molbev\/msab327"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-021-03237-4"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41564-021-00908-w"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1016\/0168-1702(84)90070-4"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrep.2021.100907"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.00474-22"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jtbi.2021.110749"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2017.11.027"},{"key":"e_1_3_4_39_2","first-page":"413","article-title":"An apparently new respiratory disease of baby chicks","volume":"78","author":"Schalk AF","year":"1931","unstructured":"Schalk AF, Hawn MC. 1931. An apparently new respiratory disease of baby chicks. J Am Vet Med Assoc 78:413\u2013422.","journal-title":"J Am Vet Med Assoc"},{"key":"e_1_3_4_40_2","first-page":"167","volume-title":"Diseases of poultry","author":"Jackwood MW","year":"2019","unstructured":"Jackwood MW, Wit S. 2019. Infectious bronchitis, p 167\u2013188. In Diseases of poultry"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virusres.2020.198002"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00140-21"},{"key":"e_1_3_4_43_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01412-08"},{"key":"e_1_3_4_44_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virusres.2014.11.021"},{"key":"e_1_3_4_45_2","doi-asserted-by":"publisher","DOI":"10.1021\/acsomega.1c00363"},{"key":"e_1_3_4_46_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1009246"},{"key":"e_1_3_4_47_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0179177"},{"key":"e_1_3_4_48_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.isci.2022.105316"},{"key":"e_1_3_4_49_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virs.2023.06.005"},{"key":"e_1_3_4_50_2","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.03176-22"},{"key":"e_1_3_4_51_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jtbi.2023.111568"},{"key":"e_1_3_4_52_2","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.27845"},{"key":"e_1_3_4_53_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.00128-22"},{"key":"e_1_3_4_54_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1608147113"},{"key":"e_1_3_4_55_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.00753-22"}],"container-title":["Journal of Virology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/jvi.00614-26","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/jvi.00614-26","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T13:04:17Z","timestamp":1787058257000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/jvi.00614-26"}},"subtitle":[],"editor":[{"given":"John","family":"Schoggins","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,8,18]]},"references-count":54,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2026,8,18]]}},"alternative-id":["10.1128\/jvi.00614-26"],"URL":"https:\/\/doi.org\/10.1128\/jvi.00614-26","relation":{},"ISSN":["0022-538X","1098-5514"],"issn-type":[{"value":"0022-538X","type":"print"},{"value":"1098-5514","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,18]]},"assertion":[{"value":"2026-04-16","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-06-10","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-07-06","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e00614-26"}}