Characterization of the interaction between outer-fiber protein VP55 of genotype III grass carp reovirus and Fibulin-4 of grass carp
Autor: | Fei Yu, Longlong Wang, Liqun Lu, Hao Sun |
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Rok vydání: | 2019 |
Předmět: |
Fish Proteins
0301 basic medicine Carps Subfamily Endogeny Aquatic Science Reoviridae Virus Fish Diseases Viral Proteins 03 medical and health sciences Genotype Animals Environmental Chemistry Extracellular Matrix Proteins biology 04 agricultural and veterinary sciences General Medicine biology.organism_classification Molecular biology Yeast Reoviridae Infections Grass carp Fibulin 030104 developmental biology 040102 fisheries 0401 agriculture forestry and fisheries Intracellular |
Zdroj: | Fish & Shellfish Immunology. 86:355-360 |
ISSN: | 1050-4648 |
Popis: | Genotype III grass carp reovirus (GCRV; representative strain, GCRV-104) belongs to the subfamily Spinareovirinae and encodes an outer-fiber protein, VP55, responsible for mediating the infection of target tissues by the virus and assisting the virus into cells. Fibulin-4/EFEMP2 protein was previously identified as a putative binding partner for VP55 in a yeast two-hybrid screening. Here, we have further characterized the association between Fibulin-4 and VP55 by using protein interaction assays. An intracellular co-localization assay showed that RFP-Fibulin-4 co-localized with GFP-VP55 in grass carp ovary (GCO) cells. Bacterially expressed GST-tagged Fibulin-4 was shown to associate with baculovirus-expressed His-tagged VP55 in a dot-blot overlay assay; moreover, baculovirus-expressed His-tagged VP55 was able to pull down GFP-Fibulin-4 expressed in the GCO cells. We performed real-time PCR and immunoblotting analysis and showed that endogenous Fibulin-4, although suppressed to a lower level in the late infection phase, is present throughout the infection course of GCRV-104 in CIK cells. In conclusion, our results indicate that grass carp Fibulin-4 interacts with VP55. The presence of Fibulin-4, a well-known secreted protein, during the infection course of GCRV-104 in grass carp cells implies its potential role during viral egression through interaction with VP55. |
Databáze: | OpenAIRE |
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