Abstract

BackgroundGrouper (Epinephelus spp) is an economically important fish species worldwide. However, viral pathogens such as nervous necrosis virus (NNV) have been causing severe infections in the fish, resulting in great loss in the grouper aquaculture industry. Yet, the understanding of the molecular mechanisms underlying the pathogenicity of NNV is still inadequate, mainly due to insufficient genomic information of the host.ResultsDe novo assembly of grouper transcriptome in the grouper kidney (GK) cells was conducted by using short read sequencing technology of Solexa/Illumina. A sum of 66,582 unigenes with mean length of 603 bp were obtained, and were annotated according to Gene Ontology (GO) and Clusters of Orthologous Groups (COG). In addition, the tag-based digital gene expression (DGE) system was used to investigate the gene expression and pathways associated with NNV infection in GK cells. The analysis revealed endoplasmic reticulum (ER) stress response was prominently affected in NNV-infected GK cells. A further analysis revealed an interaction between the NNV capsid protein and the ER chaperone immunoglobulin heavy-chain binding protein (BiP). Furthermore, exogenous expression of NNV capsid protein was able to induce XBP-1 mRNA splicing in vivo, suggesting a role of the capsid protein in the NNV-induced ER stress.ConclusionsOur data presents valuable genetic information for Epinephelus spp., which will benefit future study in this non-model but economically important species. The DGE profile of ER stress response in NNV-infected cells provides information of many important components associated with the protein processing in ER. Specifically, we showed that the viral capsid protein might play an important role in the ER stress response.

Highlights

  • Grouper (Epinephelus spp) is an economically important fish species worldwide

  • De novo sequencing and reads assembly The grouper kidney grouper kidney cell (GK cell) line was used in this study to generate a transcriptome database

  • Gene ontology (GO) and clusters of orthologous groups (COG) annotation of unigenes The GK transcriptome unigenes were annotated according to two functional annotation conventions, Gene

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Summary

Introduction

Grouper (Epinephelus spp) is an economically important fish species worldwide. Viral pathogens such as nervous necrosis virus (NNV) have been causing severe infections in the fish, resulting in great loss in the grouper aquaculture industry. Because of the exquisite quality of flesh, grouper is one of the aquaculture species with high economic value. NNV virion is non-enveloped, spherical-shaped particle with a diameter of 25–34 nm [2,6,7]. It consists of two single-stranded, positive-sense RNA genomes: RNA1 (3.1kb) encodes the viral RNAdependent RNA polymerase (RdRp) and RNA2 encodes the viral capsid protein [8]. A subgenomic RNA3 from RNA1 contains two open reading frames that encode protein B1 (111 amino acids) and B2

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