{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T13:11:32Z","timestamp":1785417092200,"version":"3.56.0"},"reference-count":52,"publisher":"American Society for Microbiology","issue":"7","license":[{"start":{"date-parts":[[2025,7,22]],"date-time":"2025-07-22T00:00:00Z","timestamp":1753142400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,7,22]],"date-time":"2025-07-22T00:00:00Z","timestamp":1753142400000},"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":["J Virol"],"published-print":{"date-parts":[[2025,7,22]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>Since the outbreak of SARS-CoV-2, viral mutations have posed significant challenges in identifying therapeutic targets and developing broad-spectrum antiviral drugs. Post-translational modifications of genes involved in interferon production and signaling pathways play a crucial role in regulating interferon responses. In this study, we employed CRISPR-Cas9 screening based on adenine base editors to investigate functional amino acids in 1,278 innate immune-related genes. This approach, which converts A-T base pairs into G-C base pairs to probe the functional importance of specific amino acids, allowed us to identify 17 vital factors involved in SARS-CoV-2 infection. Among the candidate genes, genetic knockdown of EHMT2 exhibited the strongest antiviral effect. Further analysis revealed that UNC0638, a selective inhibitor of EHMT2, significantly reduced the endosomal entry of SARS-CoV-2 in pseudovirus assays. The observed inhibitory effect was consistently observed across multiple SARS-CoV-2 variants, including Alpha, Beta, Delta, and Omicron. Mechanistically, UNC0638 reduced mature cathepsin L (CTSL) levels, impairing the proteolytic cleavage of SARS-CoV-2 spike protein and subsequent membrane fusion, a critical step for viral entry. Our findings uncover EHMT2 as a host dependency factor and reveal the antiviral mechanism of EHMT2 inhibitors through CTSL maturation blockade. These results advance the understanding of host factors in SARS-CoV-2 infection and provide a strategic framework for developing host-targeted antiviral therapies.<\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>In this study, we demonstrated that knockdown or knockout of EHMT2 inhibited SARS-CoV-2 infection, and inhibitors of EHMT2, including UNC0638, UNC0642, and BIX01294 showed similar restrictive effects. Mechanistically, the EHMT2 inhibitor UNC0638 restricts spike-mediated cell entry by inhibiting the maturation of CTSL, a critical protease required for SARS-CoV-2 entry via the endosomal pathway. Importantly, CTSL is not only essential for SARS-CoV-2 but also plays a key role in the entry of other coronaviruses that utilize similar pathways. Therefore, EHMT2 inhibitors could have broader applications as pan-coronavirus therapeutic agents.<\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/jvi.00741-25","type":"journal-article","created":{"date-parts":[[2025,6,18]],"date-time":"2025-06-18T09:00:28Z","timestamp":1750237228000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["UNC0638 inhibits SARS-CoV-2 entry by blocking cathepsin L maturation"],"prefix":"10.1128","volume":"99","author":[{"given":"Yongjun","family":"Chen","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02drdmm93","id-type":"ROR","asserted-by":"publisher"}],"name":"NHC Key Laboratory of System Biology of Pathogens, Christophe M\u00e9rieux Laboratory National Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical College","place":["Beijing, China"]},{"id":[{"id":"https:\/\/ror.org\/00see0y83","id-type":"ROR","asserted-by":"publisher"}],"name":"Key Laboratory of Pathogen Infection Prevention and Control (Peking Union Medical College) Ministry of Education","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yujin","family":"Shi","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02drdmm93","id-type":"ROR","asserted-by":"publisher"}],"name":"NHC Key Laboratory of System Biology of Pathogens, Christophe M\u00e9rieux Laboratory National Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical College","place":["Beijing, China"]},{"id":[{"id":"https:\/\/ror.org\/00see0y83","id-type":"ROR","asserted-by":"publisher"}],"name":"Key Laboratory of Pathogen Infection Prevention and Control (Peking Union Medical College) Ministry of Education","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoyan","family":"Zuo","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02drdmm93","id-type":"ROR","asserted-by":"publisher"}],"name":"NHC Key Laboratory of System Biology of Pathogens, Christophe M\u00e9rieux Laboratory National Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical College","place":["Beijing, China"]},{"id":[{"id":"https:\/\/ror.org\/00see0y83","id-type":"ROR","asserted-by":"publisher"}],"name":"Key Laboratory of Pathogen Infection Prevention and Control (Peking Union Medical College) Ministry of Education","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaojing","family":"Dong","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02drdmm93","id-type":"ROR","asserted-by":"publisher"}],"name":"NHC Key Laboratory of System Biology of Pathogens, Christophe M\u00e9rieux Laboratory National Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical College","place":["Beijing, China"]},{"id":[{"id":"https:\/\/ror.org\/00see0y83","id-type":"ROR","asserted-by":"publisher"}],"name":"Key Laboratory of Pathogen Infection Prevention and Control (Peking Union Medical College) Ministry of Education","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xia","family":"Xiao","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02drdmm93","id-type":"ROR","asserted-by":"publisher"}],"name":"NHC Key Laboratory of System Biology of Pathogens, Christophe M\u00e9rieux Laboratory National Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical College","place":["Beijing, China"]},{"id":[{"id":"https:\/\/ror.org\/00see0y83","id-type":"ROR","asserted-by":"publisher"}],"name":"Key Laboratory of Pathogen Infection Prevention and Control (Peking Union Medical College) Ministry of Education","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lan","family":"Chen","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02drdmm93","id-type":"ROR","asserted-by":"publisher"}],"name":"NHC Key Laboratory of System Biology of Pathogens, Christophe M\u00e9rieux Laboratory National Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical College","place":["Beijing, China"]},{"id":[{"id":"https:\/\/ror.org\/00see0y83","id-type":"ROR","asserted-by":"publisher"}],"name":"Key Laboratory of Pathogen Infection Prevention and Control (Peking Union Medical College) Ministry of Education","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zichun","family":"Xiang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02drdmm93","id-type":"ROR","asserted-by":"publisher"}],"name":"NHC Key Laboratory of System Biology of Pathogens, Christophe M\u00e9rieux Laboratory National Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical College","place":["Beijing, China"]},{"id":[{"id":"https:\/\/ror.org\/00see0y83","id-type":"ROR","asserted-by":"publisher"}],"name":"Key Laboratory of Pathogen Infection Prevention and Control (Peking Union Medical College) Ministry of Education","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6645-8183","authenticated-orcid":false,"given":"Lili","family":"Ren","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02drdmm93","id-type":"ROR","asserted-by":"publisher"}],"name":"NHC Key Laboratory of System Biology of Pathogens, Christophe M\u00e9rieux Laboratory National Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical College","place":["Beijing, China"]},{"id":[{"id":"https:\/\/ror.org\/00see0y83","id-type":"ROR","asserted-by":"publisher"}],"name":"Key Laboratory of Pathogen Infection Prevention and Control (Peking Union Medical College) Ministry of Education","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhuo","family":"Zhou","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02szepc22","id-type":"ROR","asserted-by":"publisher"}],"name":"State Key Laboratory of Common Mechanism Research for Major Diseases, Suzhou Institute of Systems Medicine, Chinese Academy of Medical Sciences & Peking Union Medical College","place":["Suzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8053-2423","authenticated-orcid":false,"given":"Wensheng","family":"Wei","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02v51f717","id-type":"ROR","asserted-by":"publisher"}],"name":"Biomedical Pioneering Innovation Center, Peking-Tsinghua Center for Life Sciences, Peking University Genome Editing Research Center, State Key Laboratory of Gene Function and Modulation Research, School of Life Sciences, Peking University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3455-6723","authenticated-orcid":false,"given":"Xiaobo","family":"Lei","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02drdmm93","id-type":"ROR","asserted-by":"publisher"}],"name":"NHC Key Laboratory of System Biology of Pathogens, Christophe M\u00e9rieux Laboratory National Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical College","place":["Beijing, China"]},{"id":[{"id":"https:\/\/ror.org\/00see0y83","id-type":"ROR","asserted-by":"publisher"}],"name":"Key Laboratory of Pathogen Infection Prevention and Control (Peking Union Medical College) Ministry of Education","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1116-4559","authenticated-orcid":true,"given":"Jianwei","family":"Wang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02drdmm93","id-type":"ROR","asserted-by":"publisher"}],"name":"NHC Key Laboratory of System Biology of Pathogens, Christophe M\u00e9rieux Laboratory National Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical College","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1097\/CM9.0000000000000722"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(20)30251-8"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41564-020-0695-z"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa030747"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(13)60982-4"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(20)30183-5"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-022-04594-4"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-021-04385-3"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1007\/s10787-023-01253-6"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41392-024-01956-4"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41392-023-01587-1"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2208822"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00109-21"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2020.12.005"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2023.06.005"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1002\/mco2.92"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-29896-z"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00562-20"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.15252\/embj.2020106057"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-023-06322-y"},{"key":"e_1_3_3_22_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.02085-24"},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-022-00839-1"},{"key":"e_1_3_3_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2016.06.020"},{"key":"e_1_3_3_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2012.03.022"},{"key":"e_1_3_3_26_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature13140"},{"key":"e_1_3_3_27_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2023.10.033"},{"key":"e_1_3_3_28_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature24644"},{"key":"e_1_3_3_29_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2021.01.041"},{"key":"e_1_3_3_30_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.4172"},{"key":"e_1_3_3_31_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2007.01.017"},{"key":"e_1_3_3_32_2","doi-asserted-by":"publisher","DOI":"10.7150\/jca.48310"},{"key":"e_1_3_3_33_2","doi-asserted-by":"publisher","DOI":"10.15252\/embr.202256128.32"},{"key":"e_1_3_3_34_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41420-020-00400-0"},{"key":"e_1_3_3_35_2","doi-asserted-by":"publisher","DOI":"10.1038\/nm.4257"},{"key":"e_1_3_3_36_2","doi-asserted-by":"publisher","DOI":"10.1089\/jir.2015.0006"},{"key":"e_1_3_3_37_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-021-03431-4"},{"key":"e_1_3_3_38_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1009706"},{"key":"e_1_3_3_39_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.xpro.2025.103686"},{"key":"e_1_3_3_40_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.abc6027"},{"key":"e_1_3_3_41_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.abd4585"},{"key":"e_1_3_3_42_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-17665-9"},{"key":"e_1_3_3_43_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2021.10.008"},{"key":"e_1_3_3_44_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41392-021-00800-3"},{"key":"e_1_3_3_45_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.isci.2024.110475"},{"key":"e_1_3_3_46_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-15562-9"},{"key":"e_1_3_3_47_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1008461"},{"key":"e_1_3_3_48_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41392-021-00558-8"},{"key":"e_1_3_3_49_2","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.4c11620"},{"key":"e_1_3_3_50_2","doi-asserted-by":"publisher","DOI":"10.1002\/cmdc.202100456"},{"key":"e_1_3_3_51_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41421-022-00419-w"},{"key":"e_1_3_3_52_2","doi-asserted-by":"publisher","DOI":"10.1021\/acsptsci.3c00313"},{"key":"e_1_3_3_53_2","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.24135"}],"container-title":["Journal of Virology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/jvi.00741-25","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/jvi.00741-25","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,7,22]],"date-time":"2025-07-22T13:05:33Z","timestamp":1753189533000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/jvi.00741-25"}},"subtitle":[],"editor":[{"given":"Shan-Lu","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2025,7,22]]},"references-count":52,"journal-issue":{"issue":"7","published-print":{"date-parts":[[2025,7,22]]}},"alternative-id":["10.1128\/jvi.00741-25"],"URL":"https:\/\/doi.org\/10.1128\/jvi.00741-25","relation":{},"ISSN":["0022-538X","1098-5514"],"issn-type":[{"value":"0022-538X","type":"print"},{"value":"1098-5514","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,22]]},"assertion":[{"value":"2025-04-25","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-05-25","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-06-18","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e00741-25"}}