Identification of an amino acid residue on influenza C virus M1 protein responsible for formation of the cord-like structures of the virus
| Autor: | Emi Takashita, Yoko Matsuzaki, Kanetsu Sugawara, Seiji Hongo, Hiroshi Washioka, Yasushi Muraki |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Influenzavirus C
viruses M1 protein Orthomyxoviridae Molecular Sequence Data Biology Kidney Transfection Virus Green fluorescent protein Cell Line Viral Matrix Proteins Genes Reporter Virology Complementary DNA Humans Amino Acid Sequence fungi RNA biology.organism_classification Molecular biology Microscopy Electron biology.protein Influenza C Virus Plasmids |
| Zdroj: | The Journal of general virology. 85(Pt 7) |
| ISSN: | 0022-1317 |
| Popis: | Influenza C virus-like particles (VLPs) have been generated from cloned cDNAs. A cDNA of the green fluorescent protein (GFP) gene in antisense orientation was flanked by the 5′ and 3′ non-coding regions of RNA segment 5 of the influenza C virus. The cDNA cassette was inserted between an RNA polymerase I promoter and terminator of the Pol I vector. This plasmid DNA was transfected into 293T cells together with plasmids encoding virus proteins of C/Ann Arbor/1/50 or C/Yamagata/1/88. Transfer of the supernatants of the transfected 293T cells to HMV-II cells resulted in GFP expression in the HMV-II cells. The quantification of the GFP-positive HMV-II cells indicated the presence of approximately 106 VLPs (ml supernatant)−1. Cords 50−300 μm in length were observed on transfected 293T cells, although the cords were not observed when the plasmid for M1 protein of C/Ann Arbor/1/50 was replaced with that of C/Taylor/1233/47. A series of transfection experiments with plasmids encoding M1 mutants of C/Ann Arbor/1/50 or C/Taylor/1233/47 showed that an amino acid at residue 24 of the M1 protein is responsible for cord formation. This finding provides direct evidence for a previous hypothesis that M1 protein is involved in the formation of cord-like structures protruding from the C/Yamagata/1/88-infected cells. Evidence was obtained by electron microscopy that transfected cells bearing cords produced filamentous VLPs, suggesting the potential role of the M1 protein in determining the filamentous/spherical morphology of influenza C virus. |
| Databáze: | OpenAIRE |
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