Abstract
Singapore grouper iridovirus (SGIV), belonging to genus Ranavirus, family Iridoviridae, causes great economic losses in the aquaculture industry. Previous studies demonstrated the lipid composition of intracellular unenveloped viruses, but the changes in host-cell glyceophospholipids components and the roles of key enzymes during SGIV infection still remain largely unknown. Here, the whole cell lipidomic profiling during SGIV infection was analyzed using UPLC-Q-TOF-MS/MS. The lipidomic data showed that glycerophospholipids (GPs), including phosphatidylcholine (PC), phosphatidylserine (PS), glycerophosphoinositols (PI) and fatty acids (FAs) were significantly elevated in SGIV-infected cells, indicating that SGIV infection disturbed GPs homeostasis, and then affected the metabolism of FAs, especially arachidonic acid (AA). The roles of key enzymes, such as cytosolic phospholipase A2 (cPLA2), 5-Lipoxygenase (5-LOX), and cyclooxygenase (COX) in SGIV infection were further investigated using the corresponding specific inhibitors. The inhibition of cPLA2 by AACOCF3 decreased SGIV replication, suggesting that cPLA2 might play important roles in the process of SGIV infection. Consistent with this result, the ectopic expression of EccPLA2α or knockdown significantly enhanced or suppressed viral replication in vitro, respectively. In addition, the inhibition of both 5-LOX and COX significantly suppressed SGIV replication, indicating that AA metabolism was essential for SGIV infection. Taken together, our results demonstrated for the first time that SGIV infection in vitro disturbed GPs homeostasis and cPLA2 exerted crucial roles in SGIV replication.
Highlights
Iridoviruses, members of nucleocytoplasmic large DNA viruses (NCLDV), cause great economic losses in the aquaculture industry but are emerging infectious disease agents showing a significant threat to global biodiversity [1,2]
Singapore grouper iridovirus (SGIV) infection in several host cells, such as grouper spleen (GS), GB, GK and GL evoked nonapoptotic cell death, and no apoptotic bodies were examined in infected cells [27,28]
These differential lipids belonged to 32 subclasses within the five major class, including glycerophospholipids (GPs), glycerolipids (GLs), sphingolipids (SPs), Fatty Acyls (FAs) and Sterol Lipids (STs)
Summary
Iridoviruses, members of nucleocytoplasmic large DNA viruses (NCLDV), cause great economic losses in the aquaculture industry but are emerging infectious disease agents showing a significant threat to global biodiversity [1,2]. Iridoviruses are capable of infecting invertebrates and vertebrates, such as insects, fish, amphibians, and reptiles [3]. Iridoviruses are divided into five genera: Iridovirus, Chloriridovirus, Lymphocystivirus, Megalocytivirus, and Ranavirus [3]. Similar to frog virus (FV) 3, a strain type of Ranavirus, most of the vertebrate iridovirus isolates shared the same overall replication strategy. Assembly and release of iridovirus are intimately associated with the cell membrane system, which is composed of different lipid composition. The enveloped virions enter into cells by endocytosis, whereas non-enveloped virions enter by binding the plasm membrane with the subsequent release of the viral core into the cytoplasm [4,5,6]. As with other large DNA viruses, the interaction of the internal viral membrane with either the plasma or endosomal membrane was required for release of viral DNA cores [7]. Membrane remodeling exerted crucial roles during the life cycle of iridovirus infection
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