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Using a yeast two-hybrid screen, we identified the interaction of the cytosolic tail of Murine Hepatitis Virus (MHV-CoV) M protein with Myosin Vb (MYO5B), specifically with the alternative splice variant of cellular MYO5B including exon D (MYO5B+D), which mediates interaction with Rab10. When co-expressed in human lung epithelial A549 and canine kidney epithelial MDCK cells, MYO5B+D co-localized with the MHV-CoV M protein, as well as with the M proteins from Porcine Epidemic Diarrhea Virus (PEDV-CoV), Middle East Respiratory Syndrome (MERS-CoV) and Severe Acute Respiratory Syndrome 2 (SARS-CoV-2). Co-expressed M proteins and MYO5B+D co-localized with endogenous Rab10 and Rab11a. We identified point mutations in MHV-CoV M that blocked the interaction with MYO5B+D in yeast 2-hybrid assays. One of these point mutations (E121K) was previously shown to block MHV-CoV virion assembly and its interaction with MYO5B+D. The E to K mutation at homologous positions in PEDV-CoV, MERS-CoV and SARS-CoV-2 M proteins also blocked colocalization with MYO5B+D. The knockdown of Rab10 blocked the co-localization of M proteins with MYO5B+D and was rescued by re-expression of CFP-Rab10. Our results suggest that CoV M proteins traffic through Rab10-containing systems, in association with MYO5B+D.<\/jats:p>","DOI":"10.3390\/cells13020126","type":"journal-article","created":{"date-parts":[[2024,1,10]],"date-time":"2024-01-10T05:47:21Z","timestamp":1704865641000},"page":"126","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Coronavirus M Protein Trafficking in Epithelial Cells Utilizes a Myosin Vb Splice Variant and Rab10"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7163-0830","authenticated-orcid":false,"given":"Lynne A.","family":"Lapierre","sequence":"first","affiliation":[{"name":"Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN 37232, USA"},{"name":"Epithelial Biology Center, Vanderbilt University School of Medicine, Nashville, TN 37232, USA"},{"name":"Nashville VA Medical Center, Nashville, TN 37212, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph T.","family":"Roland","sequence":"additional","affiliation":[{"name":"Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN 37232, USA"},{"name":"Epithelial Biology Center, Vanderbilt University School of Medicine, Nashville, TN 37232, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elizabeth H.","family":"Manning","sequence":"additional","affiliation":[{"name":"Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN 37232, USA"},{"name":"Epithelial Biology Center, Vanderbilt University School of Medicine, Nashville, TN 37232, USA"},{"name":"Nashville VA Medical Center, Nashville, TN 37212, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Catherine","family":"Caldwell","sequence":"additional","affiliation":[{"name":"Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN 37232, USA"},{"name":"Epithelial Biology Center, Vanderbilt University School of Medicine, Nashville, TN 37232, USA"},{"name":"Nashville VA Medical Center, Nashville, TN 37212, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Honor L.","family":"Glenn","sequence":"additional","affiliation":[{"name":"Biodesign Institute Center for Immunotherapy, Vaccines & Virotherapy, Tempe, AZ 85287, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pierre-Olivier","family":"Vidalain","sequence":"additional","affiliation":[{"name":"Equipe Infections Virales, M\u00e9tabolisme et Immunit\u00e9, Centre International de Recherche en Infectiologie (CIRI), Univ. Lyon, INSERM U1111, CNRS UMR5308, Ecole Normale Sup\u00e9rieure de Lyon, Universit\u00e9 Claude Bernard Lyon 1, 69008 Lyon, France"},{"name":"Unit\u00e9 G\u00e9nomique Virale et Vaccination, Institut Pasteur, CNRS UMR3569, 75015 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5937-5654","authenticated-orcid":false,"given":"Frederic","family":"Tangy","sequence":"additional","affiliation":[{"name":"Viral Genomics and Vaccination Unit, Department of Virology, Institut Pasteur, CNRS UMR3569, 75015 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4545-9208","authenticated-orcid":false,"given":"Brenda G.","family":"Hogue","sequence":"additional","affiliation":[{"name":"Biodesign Institute Center for Immunotherapy, Vaccines & Virotherapy, Tempe, AZ 85287, USA"},{"name":"Center for Applied Structural Discovery, Biodesign Institute, Tempe, AZ 85287, USA"},{"name":"School of Life Sciences, Arizona State University, Phoenix, AZ 85004, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4459-9874","authenticated-orcid":false,"given":"C. 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M.","family":"de Haan","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine, Department of Biomolecular Health Sciences, Division of Infectious Diseases and Immunology, Section Virology, University of Utrecht, 3584 CS Utrecht, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7833-2940","authenticated-orcid":false,"given":"James R.","family":"Goldenring","sequence":"additional","affiliation":[{"name":"Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN 37232, USA"},{"name":"Epithelial Biology Center, Vanderbilt University School of Medicine, Nashville, TN 37232, USA"},{"name":"Nashville VA Medical Center, Nashville, TN 37212, USA"},{"name":"Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,1,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1038\/s41579-018-0118-9","article-title":"Origin and evolution of pathogenic coronaviruses","volume":"17","author":"Cui","year":"2019","journal-title":"Nat. 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