{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,10]],"date-time":"2026-06-10T14:26:07Z","timestamp":1781101567537,"version":"3.54.1"},"reference-count":58,"publisher":"National Academy of Sciences","issue":"49","license":[{"start":{"date-parts":[[2025,12,4]],"date-time":"2025-12-04T00:00:00Z","timestamp":1764806400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"the National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["31672534"],"award-info":[{"award-number":["31672534"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100017596","name":"\u9655\u897f\u7701\u79d1\u5b66\u6280\u672f\u5385 | Natural Science Basic Research Program of Shaanxi Province","doi-asserted-by":"publisher","award":["2024JC-JCQN-24"],"award-info":[{"award-number":["2024JC-JCQN-24"]}],"id":[{"id":"10.13039\/501100017596","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Northwest A&F University","award":["Z10202190601"],"award-info":[{"award-number":["Z10202190601"]}]}],"content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2025,12,9]]},"abstract":"<jats:p>Hepatitis E virus (HEV), the causative agent of hepatitis E, is threatening public health globally. Due to the shortage of efficient in vitro cell culture systems and in vivo model, the viral replication and pathogenesis mechanisms remain largely unknown. Here, we found that HEV-ORF3 protein showed prion-like properties in HEV-infected cells and existed as both monomer and SDS-resistant aggregate forms. In an in vitro cell-free model, incubation of ORF3 monomer with its aggregates could effectively convert ORF3 monomer into aggregates, mirroring a typical characteristic of prion. In addition, the prion domain (PrD) of a classic yeast prion Sup35 could be functionally replaced by full-length HEV-ORF3 or its N-terminal candidate PrD (cPrD). An F10S substitution in the ORF3-cPrD impaired HEV-ORF3 aggregation propensity and blocked the function of ORF3 in enhancing the stability of microtubules in HEV-infected cells, thus led to the inhibition of viral capsid translocation to microtubules and virion release from infected cells. In Mongolian gerbil models, HEV bearing ORF3F10S mutation demonstrated attenuated virulence in vivo compared with wild-type HEV, as evidenced by reduced viremia and viral shedding, as well as alleviated pathological changes of liver tissue in gerbils infected by HEV-ORF3F10S mutant. In conclusion, our data suggest that HEV-ORF3 is a prion-like protein which is involved in viral capsid translocation and virion release, supporting the hypothesis that the self-propagating properties of prion proteins or prion-like proteins are widely exploited in nature and play diverse roles in physiological function.<\/jats:p>","DOI":"10.1073\/pnas.2511801122","type":"journal-article","created":{"date-parts":[[2025,12,4]],"date-time":"2025-12-04T19:11:27Z","timestamp":1764875487000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":1,"title":["The prion-like characteristic of ORF3 contributes to virion release and pathogenesis of hepatitis E virus"],"prefix":"10.1073","volume":"122","author":[{"given":"Yajing","family":"Wang","sequence":"first","affiliation":[{"name":"Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University","place":["Yangling, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-6386-7793","authenticated-orcid":false,"given":"Houlu","family":"Tian","sequence":"additional","affiliation":[{"name":"Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University","place":["Yangling, China"]},{"name":"Department of Bioengineering, College of Life Sciences, Northwest A&F University","place":["Yangling, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Na","family":"Shi","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, College of Life Sciences, Northwest A&F University","place":["Yangling, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunyan","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University","place":["Yangling, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yaowei","family":"Huang","sequence":"additional","affiliation":[{"name":"Guangdong Laboratory for Lingnan Modern Agriculture, College of Veterinary Medicine, South China Agricultural University","place":["Guangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3162-4070","authenticated-orcid":false,"given":"Yongling","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Infectious Diseases, The Affiliated Taizhou People\u2019s Hospital of Nanjing Medical University","place":["Taizhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6226-869X","authenticated-orcid":false,"given":"Qiang","family":"Ding","sequence":"additional","affiliation":[{"name":"Center for Infectious Disease Research, School of Medicine, Tsinghua University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xu","family":"Zheng","sequence":"additional","affiliation":[{"name":"Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University","place":["Yangling, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qin","family":"Zhao","sequence":"additional","affiliation":[{"name":"Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University","place":["Yangling, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8091-7630","authenticated-orcid":false,"given":"Zhaonong","family":"Hu","sequence":"additional","affiliation":[{"name":"Institute of Pesticide Science, College of Plant Protection, Northwest A&F University","place":["Yangling, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiaojiao","family":"Luo","sequence":"additional","affiliation":[{"name":"Institute of Pesticide Science, College of Plant Protection, Northwest A&F University","place":["Yangling, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lele","family":"Feng","sequence":"additional","affiliation":[{"name":"Institute of Pesticide Science, College of Plant Protection, Northwest A&F University","place":["Yangling, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lingdong","family":"Xu","sequence":"additional","affiliation":[{"name":"Laboratory Animal Center, Zhejiang University","place":["Hangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5214-540X","authenticated-orcid":false,"given":"Mick F.","family":"Tuite","sequence":"additional","affiliation":[{"name":"Kent Fungal Group, Division of Natural Sciences, School of Biosciences, University of Kent","place":["Canterbury, United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8258-0683","authenticated-orcid":false,"given":"Hongying","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, College of Life Sciences, Northwest A&F University","place":["Yangling, China"]},{"name":"Institute of Future Agriculture, Northwest A&F University","place":["Yangling, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4442-8004","authenticated-orcid":false,"given":"Yuchen","family":"Nan","sequence":"additional","affiliation":[{"name":"Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University","place":["Yangling, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2025,12,4]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00822-17"},{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2017.02396"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1055\/s-0033-1338113"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrneurol.2015.234"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1005521"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.89.17.8259"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00046-06"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1614955114"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02259-06"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02571-08"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1055\/s-0033-1338110"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.71.12.9045-9053.1997"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1099\/vir.0.010561-0"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1099\/vir.0.035378-0"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-08180-z"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1604478113"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.aai7776"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2418751122"},{"key":"e_1_3_4_19_2","first-page":"838","article-title":"Screening for amyloid aggregation by semi-denaturing detergent-agarose gel electrophoresis","volume":"17","author":"Halfmann R.","year":"2008","unstructured":"R. Halfmann, S. Lindquist, Screening for amyloid aggregation by semi-denaturing detergent-agarose gel electrophoresis. JoVE 17, 838 (2008).","journal-title":"JoVE"},{"key":"e_1_3_4_20_2","first-page":"39","article-title":"Amyloids, Congo red and the apple-green effect","volume":"66","author":"Jagusiak A.","year":"2019","unstructured":"A. Jagusiak , Amyloids, Congo red and the apple-green effect. Acta Biochim. Pol. 66, 39\u201346 (2019).","journal-title":"Acta Biochim. Pol."},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2011.06.041"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2009.02.044"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.7754373"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1007\/s002840010251"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.3390\/ijms23105738"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btu310"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1721345115"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00705-013-1912-3"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-virology-031413-085359"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1007471"},{"key":"e_1_3_4_31_2","first-page":"50031","article-title":"Detection of protein palmitoylation in cultured hippocampal neurons by immunoprecipitation and acyl-biotin exchange (ABE)","volume":"72","author":"Brigidi G. S.","year":"2013","unstructured":"G. S. Brigidi, S. X. Bamji, Detection of protein palmitoylation in cultured hippocampal neurons by immunoprecipitation and acyl-biotin exchange (ABE). JoVE 72, 50031 (2013).","journal-title":"JoVE"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2010.11.023"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-30376-y"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M200185200"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2017.09.020"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.02193-21"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1042\/BSR20181415"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1093\/labmed\/lmab022"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-019-55528-6"},{"key":"e_1_3_4_40_2","doi-asserted-by":"publisher","DOI":"10.3109\/13506129.2010.527448"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.6815801"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41576-018-0011-4"},{"key":"e_1_3_4_43_2","doi-asserted-by":"publisher","DOI":"10.1038\/ncb1843"},{"key":"e_1_3_4_44_2","doi-asserted-by":"publisher","DOI":"10.1038\/srep45720"},{"key":"e_1_3_4_45_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2017.02239"},{"key":"e_1_3_4_46_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-27256-w"},{"key":"e_1_3_4_47_2","doi-asserted-by":"publisher","DOI":"10.3390\/epigenomes6010004"},{"key":"e_1_3_4_48_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2012.01.004"},{"key":"e_1_3_4_49_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1067484"},{"key":"e_1_3_4_50_2","doi-asserted-by":"publisher","DOI":"10.1128\/EC.00068-09"},{"key":"e_1_3_4_51_2","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-82-11-2747"},{"key":"e_1_3_4_52_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.78.15.7990-8001.2004"},{"key":"e_1_3_4_53_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2009.03.041"},{"key":"e_1_3_4_54_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01717-07"},{"key":"e_1_3_4_55_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00538-17"},{"key":"e_1_3_4_56_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02606-10"},{"key":"e_1_3_4_57_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00488-16"},{"key":"e_1_3_4_58_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-11015-4"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.2511801122","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,4]],"date-time":"2025-12-04T19:11:42Z","timestamp":1764875502000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/10.1073\/pnas.2511801122"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,4]]},"references-count":58,"journal-issue":{"issue":"49","published-print":{"date-parts":[[2025,12,9]]}},"alternative-id":["10.1073\/pnas.2511801122"],"URL":"https:\/\/doi.org\/10.1073\/pnas.2511801122","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,4]]},"assertion":[{"value":"2025-05-23","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-10-31","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-12-04","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e2511801122"}}