{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,14]],"date-time":"2026-07-14T01:51:36Z","timestamp":1783993896906,"version":"3.55.0"},"reference-count":61,"publisher":"American Society for Microbiology","issue":"10","license":[{"start":{"date-parts":[[2024,10,16]],"date-time":"2024-10-16T00:00:00Z","timestamp":1729036800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2024,10,16]],"date-time":"2024-10-16T00:00:00Z","timestamp":1729036800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"crossref","award":["401821119\/GRK2504"],"award-info":[{"award-number":["401821119\/GRK2504"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Bayerische Forschungsstiftung, non-validated organization name","award":["DeeP-CMV\/AP-A"],"award-info":[{"award-number":["DeeP-CMV\/AP-A"]}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["mBio"],"published-print":{"date-parts":[[2024,10,16]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:sec>\n            <jats:title\/>\n            <jats:p>Cell entry is a crucial step for a virus to infect a host cell. Human cytomegalovirus utilizes glycoprotein B (gB) to fuse the viral and host cell membranes upon receptor binding of gH\/gL-containing complexes. Fusion is mediated by major conformational changes of gB from a metastable pre-fusion to a stable post-fusion state whereby the central trimeric coiled-coils, formed by domain (Dom)III \u03b1 helices, remain structurally nearly unchanged. To better understand the role of the stable core, we individually introduced three potentially helix-breaking or one disulfide bond-breaking mutation in the DIII \u03b13 to study different aspects of the viral behavior upon long-term culturing. Two of the three helix-breaking mutations, gB_Y494P and gB_I495P, were lethal for the virus in either fibroblasts or epithelial cells. The third substitution, gB_G493P, on the other hand, displayed a delayed replication and spread, which was more pronounced in epithelial cells, hinting at an impaired fusion. Interestingly, the disulfide bond-breaker mutation, gB_C507S, performed strikingly differently in the two cell types - lethal in epithelial cells and an atypical phenotype in fibroblasts, respectively. Replication curve analyses paired with the infection efficiency, the spread morphology, and the cell\u2013cell fusogenicity suggest a dysregulated fusion process, which could be reverted by second-site mutations mapping predominantly to gB DomV. Our findings underline the functional importance of a stable DomIII core for a well-regulated DomV rearrangement during fusion.<\/jats:p>\n            <jats:sec>\n              <jats:title>IMPORTANCE<\/jats:title>\n              <jats:p>Human cytomegalovirus (HCMV) can establish a lifelong infection. In most people, the infection follows an asymptomatic course; however, it is a major cause of morbidity and mortality in immunocompromised patients or neonates. HCMV has a very broad cell tropism, ranging from fibroblasts to epi- and endothelial cells. The virus uses different entry pathways utilizing the core fusion machinery consisting of glycoprotein complexes gH\/gL and glycoprotein B (gB). The fusion protein gB undergoes fundamental rearrangements from a metastable pre-fusion to a stable post-fusion conformation. Here, we characterized the viral behavior after the introduction of four single-point mutations in the gB central core. These led to various cell type-specific atypical phenotypes and the emergence of compensatory mutations, demonstrating an important interaction between domains III and V. We provide a new basis for the development of a structurally and functionally altered gB, which can further serve as a tool for drug and vaccine development.<\/jats:p>\n            <\/jats:sec>\n          <\/jats:sec>","DOI":"10.1128\/mbio.01812-24","type":"journal-article","created":{"date-parts":[[2024,9,24]],"date-time":"2024-09-24T13:09:14Z","timestamp":1727183354000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":3,"title":["Growth defect of domain III glycoprotein B mutants of human cytomegalovirus reverted by compensatory mutations co-localizing in post-fusion conformation"],"prefix":"10.1128","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-2949-1378","authenticated-orcid":false,"given":"Madlen","family":"Mollik","sequence":"first","affiliation":[{"name":"Center for Virology, Medical University of Vienna, Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas","family":"Rohorzka","sequence":"additional","affiliation":[{"name":"Center for Virology, Medical University of Vienna, Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaohan","family":"Chen","sequence":"additional","affiliation":[{"name":"Virologisches Institut, Klinische und Molekulare Virologie, Friedrich Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg, Erlangen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara","family":"Kropff","sequence":"additional","affiliation":[{"name":"Virologisches Institut, Klinische und Molekulare Virologie, Friedrich Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg, Erlangen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lukas","family":"Eisler","sequence":"additional","affiliation":[{"name":"Center for Virology, Medical University of Vienna, Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"B\u00fcsra","family":"K\u00fclekci","sequence":"additional","affiliation":[{"name":"Center for Virology, Medical University of Vienna, Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0673-8335","authenticated-orcid":true,"given":"Elisabeth","family":"Puchhammer-St\u00f6ckl","sequence":"additional","affiliation":[{"name":"Center for Virology, Medical University of Vienna, Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4820-2712","authenticated-orcid":false,"given":"Marco","family":"Thomas","sequence":"additional","affiliation":[{"name":"Virologisches Institut, Klinische und Molekulare Virologie, Friedrich Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg, Erlangen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1936-0404","authenticated-orcid":false,"given":"Irene","family":"G\u00f6rzer","sequence":"additional","affiliation":[{"name":"Center for Virology, Medical University of Vienna, Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.3390\/v14081816"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-020-00448-w"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.76.12.6073-6082.2002"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2017.00934"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1005227"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abf3178"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1007452"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01623-08"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01276-20"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.3390\/v14020296"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1424818112"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2021.01.036"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/0042-6822(92)90200-9"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.80.2.710-722.2006"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-80-11-2867"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-0716-1111-1_1"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1189\/jlb.0112040"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1099\/vir.0.018994-0"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI42955"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.3390\/v13040614"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.3390\/v13091780"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006273"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.78.3.1289-1300.2004"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.3390\/v13061106"},{"key":"e_1_3_4_26_2","first-page":"1","article-title":"Epidemiology of human cytomegalovirus strains through comparison of methodological approaches to explore gN variants","volume":"32","author":"Pignatelli S","year":"2009","unstructured":"Pignatelli S, Dal Monte P. 2009. Epidemiology of human cytomegalovirus strains through comparison of methodological approaches to explore gN variants. New Microbiol 32:1\u201310.","journal-title":"New Microbiol"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.2217\/fvl.10.87"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.20235"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00578-15"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/jiz208"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1907447116"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM200105033441804"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02051-19"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.3390\/v14071508"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00606-18"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01325-15"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00107-20"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abc1726"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-43011-w"},{"key":"e_1_3_4_40_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1001256"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2017.01609"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00104-12"},{"key":"e_1_3_4_43_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01948-18"},{"key":"e_1_3_4_44_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00557-21"},{"key":"e_1_3_4_45_2","doi-asserted-by":"publisher","DOI":"10.3390\/v15071500"},{"key":"e_1_3_4_46_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.coviro.2012.01.001"},{"key":"e_1_3_4_47_2","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.01864-22"},{"key":"e_1_3_4_48_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-36683-x"},{"key":"e_1_3_4_49_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00119-11"},{"key":"e_1_3_4_50_2","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abm2536"},{"key":"e_1_3_4_51_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00612-21"},{"key":"e_1_3_4_52_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02249-09"},{"key":"e_1_3_4_53_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006281"},{"key":"e_1_3_4_54_2","doi-asserted-by":"publisher","DOI":"10.1099\/jgv.0.000475"},{"key":"e_1_3_4_55_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00034-20"},{"key":"e_1_3_4_56_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0509201102"},{"key":"e_1_3_4_57_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.79.16.10330-10338.2005"},{"key":"e_1_3_4_58_2","doi-asserted-by":"publisher","DOI":"10.1099\/vir.0.81921-0"},{"key":"e_1_3_4_59_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.05354-11"},{"key":"e_1_3_4_60_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-60761-652-8_30"},{"key":"e_1_3_4_61_2","doi-asserted-by":"publisher","DOI":"10.1093\/ve\/veac076"},{"key":"e_1_3_4_62_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12864-021-08272-z"}],"container-title":["mBio"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/mbio.01812-24","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/mbio.01812-24","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,10,16]],"date-time":"2024-10-16T13:11:39Z","timestamp":1729084299000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/mbio.01812-24"}},"subtitle":[],"editor":[{"given":"Anne","family":"Moscona","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2024,10,16]]},"references-count":61,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2024,10,16]]}},"alternative-id":["10.1128\/mbio.01812-24"],"URL":"https:\/\/doi.org\/10.1128\/mbio.01812-24","relation":{},"ISSN":["2150-7511"],"issn-type":[{"value":"2150-7511","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,10,16]]},"assertion":[{"value":"2024-06-19","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-08-29","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-09-24","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e01812-24"}}