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Interaction between Two Iridovirus Core Proteins and Their Effects on Ranavirus (RGV) Replication in Cells from Different Species

1
State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Wuhan 430072, China
2
The Innovation Academy of Seed Design, Chinese Academy of Sciences, Beijing 100101, China
*
Author to whom correspondence should be addressed.
Viruses 2019, 11(5), 416; https://doi.org/10.3390/v11050416
Received: 1 March 2019 / Revised: 28 April 2019 / Accepted: 30 April 2019 / Published: 4 May 2019
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Abstract

The two putative proteins RGV-63R and RGV-91R encoded by Rana grylio virus (RGV) are DNA polymerase and proliferating cell nuclear antigen (PCNA) respectively, and are core proteins of iridoviruses. Here, the interaction between RGV-63R and RGV-91R was detected by a yeast two-hybrid (Y2H) assay and further confirmed by co-immunoprecipitation (co-IP) assays. Subsequently, RGV-63R or RGV-91R were expressed alone or co-expressed in two kinds of aquatic animal cells including amphibian Chinese giant salamander thymus cells (GSTCs) and fish Epithelioma papulosum cyprinid cells (EPCs) to investigate their localizations and effects on RGV genome replication. The results showed that their localizations in the two kinds of cells are consistent. RGV-63R localized in the cytoplasm, while RGV-91R localized in the nucleus. However, when co-expressed, RGV-63R localized in both the cytoplasm and the nucleus, and colocalized with RGV-91R in the nucleus. 91R△NLS represents the RGV-91R deleting nuclear localization signal, which is localized in the cytoplasm and colocalized with RGV-63R in the cytoplasm. qPCR analysis revealed that sole expression and co-expression of the two proteins in the cells of two species significantly promoted RGV genome replication, while varying degrees of viral genome replication levels may be linked to the cell types. This study provides novel molecular evidence for ranavirus cross-species infection and replication. View Full-Text
Keywords: Rana grylio virus (RGV); iridovirus core proteins; protein interaction; aquatic animals; cross-species transmission; yeast two-hybrid (Y2H); co-immunoprecipitation (Co-IP) Rana grylio virus (RGV); iridovirus core proteins; protein interaction; aquatic animals; cross-species transmission; yeast two-hybrid (Y2H); co-immunoprecipitation (Co-IP)
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).
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Zeng, X.-T.; Zhang, Q.-Y. Interaction between Two Iridovirus Core Proteins and Their Effects on Ranavirus (RGV) Replication in Cells from Different Species. Viruses 2019, 11, 416.

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