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.
Highlights
A large number of aquatic viruses regulate population dynamics and community interactions in aquatic ecosystems [1]
The interactions of Rana grylio virus (RGV)-63R with other iridoviral core proteins from RGV were tested by a Y2H assay, showing that RGV-63R interacts with RGV-91R
It is possible that the interaction between RGV-63R and RGV-91R in yeast is weak and only activates the reporter gene HIS3, but does not activate the reporter expressed alone in giant salamander thymus cells (GSTCs) (Figure 5), which was in line with expectations because of the long-term preservation of RGV in Epithelioma papulosum cyprinid cells (EPCs), not in GSTCs
Summary
A large number of aquatic viruses regulate population dynamics and community interactions in aquatic ecosystems [1]. They are involved in aquatic animal diseases. Aquatic animal viruses often infect shellfishes, fishes, amphibians, reptiles and aquatic mammals [2,3,4,5]. Ranaviruses (Rana grylio virus, RGV) are members of the family Iridoviridae that are large, double-stranded DNA viruses and infect ectothermic vertebrates [6]. Ranaviruses are capable of crossing species barriers of numerous ectothermic vertebrates and can spread between different species [7,8,9]. Science and technology have been applied to a wide range of studies on aquatic viruses [11]
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