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Transfection of different cell types with FCoV 3b fused to enhanced green fluorescent protein (EGFP) or 3\u00d7FLAG confirmed localization of FCoV 3b in the mitochondria and nucleoli. Using serial truncated mutants, we showed that nucleolar accumulation is controlled by a joint nucleolar and nuclear localization signal (NoLS\/NLS) in which the identified overlapping pat4 motifs (residues 53\u201357) play a critical role. Mutational analysis also revealed that mitochondrial translocation is mediated by N-terminal residues 10\u201335, in which a Tom20 recognition motif (residues 13\u201317) and two other overlapping hexamers (residues 24\u201330) associated with mitochondrial targeting were identified. In addition, a second Tom20 recognition motif was identified further downstream (residues 61\u201365), although the mitochondrial translocation evoked by these residues seemed less efficient as a diffuse cytoplasmic distribution was also observed. Assessing the spatiotemporal distribution of FCoV 3b did not provide convincing evidence of dynamic shuttling behaviour between the nucleoli and the mitochondria.<\/jats:p>","DOI":"10.1099\/jgv.0.001321","type":"journal-article","created":{"date-parts":[[2019,9,24]],"date-time":"2019-09-24T10:13:07Z","timestamp":1569319987000},"page":"1417-1430","source":"Crossref","is-referenced-by-count":5,"title":["Identification of peptide domains involved in the subcellular localization of the feline coronavirus 3b protein"],"prefix":"10.1099","volume":"100","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0967-1831","authenticated-orcid":false,"given":"Delphine D.","family":"Acar","sequence":"first","affiliation":[{"name":"Department of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7150-7015","authenticated-orcid":false,"given":"Veerle J. 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