Identification of peptide domains involved in the subcellular localization of the feline coronavirus 3b protein
| Autor: | Herman W. Favoreel, Veerle J. E. Stroobants, Xavier Saelens, Hans Nauwynck, Delphine D. Acar |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Feline coronavirus 040301 veterinary sciences Nucleolus Nuclear Localization Signals Viral Nonstructural Proteins Biology medicine.disease_cause Feline Infectious Peritonitis Green fluorescent protein 0403 veterinary science Open Reading Frames 03 medical and health sciences Protein Domains Virology medicine Animals Coronavirus Feline 04 agricultural and veterinary sciences Subcellular localization Feline infectious peritonitis Mitochondria Cell biology Protein Transport Open reading frame 030104 developmental biology Cytoplasm Cats Cell Nucleolus Nuclear localization sequence Research Article |
| Zdroj: | Journal of General Virology. 100:1417-1430 |
| ISSN: | 1465-2099 0022-1317 |
| DOI: | 10.1099/jgv.0.001321 |
| Popis: | Feline coronavirus (FCoV) has been identified as the aetiological agent of feline infectious peritonitis (FIP), a highly fatal systemic disease in cats. FCoV open reading frame 3 (ORF3) encodes accessory proteins 3a, 3b and 3 c. The FCoV 3b accessory protein consists of 72 amino acid residues and localizes to nucleoli and mitochondria. The present work focused on peptide domains within FCoV 3b that drive its intracellular trafficking. Transfection of different cell types with FCoV 3b fused to enhanced green fluorescent protein (EGFP) or 3×FLAG 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 53–57) play a critical role. Mutational analysis also revealed that mitochondrial translocation is mediated by N-terminal residues 10–35, in which a Tom20 recognition motif (residues 13–17) and two other overlapping hexamers (residues 24–30) associated with mitochondrial targeting were identified. In addition, a second Tom20 recognition motif was identified further downstream (residues 61–65), 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. |
| Databáze: | OpenAIRE |
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