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Like other coronaviruses before, SARS-CoV-2 likely emerged after its transmission from bats. The successful propagation of SARS-CoV-2 in humans might have been facilitated by usurping evolutionarily conserved cellular factors to execute crucial steps in its life cycle, such as the generation of replication organelles\u2014membrane structures where coronaviruses assemble their replication-transcription complex. In this study, we found that RAB5, which is highly conserved across mammals, is a critical host dependency factor for the replication of the SARS-CoV-2 genome. Our results also suggest that SARS-CoV-2 uses RAB5\n              <jats:sup>+<\/jats:sup>\n              membranes to build replication organelles with the aid of COPB1, a component of the COP-I complex, and that the virus protein NSP6 participates in this process. Hence, targeting NSP6 represents a promising approach to interfere with SARS-CoV-2 RNA synthesis and halt its propagation.\n            <\/jats:p>\n            <jats:sec>\n              <jats:title>IMPORTANCE<\/jats:title>\n              <jats:p>In this study, we sought to identify the host dependency factors that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) uses for the generation of replication organelles: cellular membranous structures that SARS-CoV-2 builds in order to support the replication and transcription of its genome. We uncovered that RAB5 is an important dependency factor for SARS-CoV-2 replication and the generation of replication organelles, and that the viral protein NSP6 participates in this process. Hence, NSP6 represents a promising target to halt SARS-CoV-2 replication.<\/jats:p>\n            <\/jats:sec>\n          <\/jats:sec>","DOI":"10.1128\/mbio.03314-24","type":"journal-article","created":{"date-parts":[[2025,4,1]],"date-time":"2025-04-01T13:02:49Z","timestamp":1743512569000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":5,"title":["RAB5 is a host dependency factor for the generation of SARS-CoV-2 replication organelles"],"prefix":"10.1128","volume":"16","author":[{"given":"Yuexuan","family":"Chen","sequence":"first","affiliation":[{"name":"Microbiology and Immunology, University of Rochester Medical Center","place":["Rochester, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susanne","family":"Klute","sequence":"additional","affiliation":[{"name":"Institute of Molecular Virology, Ulm University Medical Center","place":["Ulm, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Konstantin Maria Johannes","family":"Sparrer","sequence":"additional","affiliation":[{"name":"Institute of Molecular Virology, Ulm University Medical Center","place":["Ulm, Germany"]},{"name":"German Center for Neurodegenerative Diseases (DZNE)","place":["Ulm, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1923-6414","authenticated-orcid":true,"given":"Ruth","family":"Serra-Moreno","sequence":"additional","affiliation":[{"name":"Microbiology and Immunology, University of Rochester Medical Center","place":["Rochester, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tmaid.2020.101830"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-021-00652-2"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-17687-3"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12985-015-0422-1"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/S2542-5196(21)00258-8"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1002\/jgm.794"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1080\/13102818.2020.1843539"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/nm1151"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00524-13"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-022-04835-6"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.3000715"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tim.2020.05.009"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01658-17"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-022-04469-x"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1004166"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.antiviral.2023.105590"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1080\/22221751.2023.2209208"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro1890"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1006\/viro.1997.8906"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.75.23.11664-11676.2001"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02501-05"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2006.11.027"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.0030156"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-642-00302-8_16"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1126766"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.0060226"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.4161\/auto.7.11.16642"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M306124200"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2010.05.013"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2021.110049"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2020.11.003"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.3390\/ijms24010720"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-020-03745-y"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1034\/j.1600-0854.2000.010902.x"},{"key":"e_1_3_4_36_2","first-page":"171","article-title":"Effectors of GTPase Rab5 in endocytosis and signal transduction","volume":"55","author":"Olchowik M","year":"2009","unstructured":"Olchowik M, Miaczy\u0144ska M. 2009. 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