Iridovirus poses a substantial threat to global aquaculture due to its high mortality rate;however,the molecular mechanisms underpinning its pathogenesis are not well elucidated.Here,a multi-omics approach was applied...Iridovirus poses a substantial threat to global aquaculture due to its high mortality rate;however,the molecular mechanisms underpinning its pathogenesis are not well elucidated.Here,a multi-omics approach was applied to groupers infected with Singapore grouper iridovirus(SGIV),focusing on the roles of key metabolites.Results showed that SGIV induced obvious histopathological damage and changes in metabolic enzymes within the liver.Furthermore,SGIV significantly reduced the contents of lipid droplets,triglycerides,cholesterol,and lipoproteins.Metabolomic analysis indicated that the altered metabolites were enriched in 19 pathways,with a notable down-regulation of lipid metabolites such as glycerophosphates and alpha-linolenic acid(ALA),consistent with disturbed lipid homeostasis in the liver.Integration of transcriptomic and metabolomic data revealed that the top enriched pathways were related to cell growth and death and nucleotide,carbohydrate,amino acid,and lipid metabolism,supporting the conclusion that SGIV infection induced liver metabolic reprogramming.Further integrative transcriptomic and proteomic analysis indicated that SGIV infection activated crucial molecular events in a phagosome-immune depression-metabolism dysregulation-necrosis signaling cascade.Of note,integrative multi-omics analysis demonstrated the consumption of ALA and linoleic acid(LA)metabolites,and the accumulation of L-glutamic acid(GA),accompanied by alterations in immune,inflammation,and cell death-related genes.Further experimental data showed that ALA,but not GA,suppressed SGIV replication by activating antioxidant and anti-inflammatory responses in the host.Collectively,these findings provide a comprehensive resource for understanding host response dynamics during fish iridovirus infection and highlight the antiviral potential of ALA in the prevention and treatment of iridoviral diseases.展开更多
Virus infection consists of entry, synthesis of macro- molecular components, virus assembly and release. Understanding of the mechanisms underlying each event is necessary for the intervention of virus infection in hu...Virus infection consists of entry, synthesis of macro- molecular components, virus assembly and release. Understanding of the mechanisms underlying each event is necessary for the intervention of virus infection in human healthcare and agriculture. Here we report the visualization of Singapore grouper iridovirus （SGIV） assembly in the medaka haploid embryonic stem （ES） cell line HX1. SGIV is a highly infectious DNA virus that causes a massive loss in marine aquaculture. Ectopic expression of VP88GFP, a fusion between green fluo- rescent protein and the envelope protein VP088, did not compromise the ES cell properties and susceptibility to SGIV infection. Although VP88GFP disperses evenly in the cytoplasm of non-infected cells, it undergoes aggregation and redistribution in SGIV-infected cells. Real-time visualization revealed multiple key stages of VP88GFP redistribution and the dynamics of viral assembly site （VAS）. Specifically, VP88GFP entry into and condensation in the VAS occurred within a 6-h duration, a similar duration was observed also for the release of VP88GFP-containing SGIV out of the cell, Taken together, VP088 is an excellent marker for visu- alizing the SGIV infection process. Our results provide new insight into macromolecular component recruit- ment and SGIV assembly.展开更多
Myeloid differentiation factor 88(MyD88) is a universal adaptor protein involved in Toll-like receptors and in interleukin-1 receptor-induced nuclear factor-kappaB(NF-κB) activation.In this study,a new MyD88 gene(des...Myeloid differentiation factor 88(MyD88) is a universal adaptor protein involved in Toll-like receptors and in interleukin-1 receptor-induced nuclear factor-kappaB(NF-κB) activation.In this study,a new MyD88 gene(designated as Og-MyD88) was cloned from orange-spotted grouper,Epinephelus coioides,based on the expressed sequence tag(EST) obtained following Roche 454 GS-FLX sequencing.The full-length Og-MyD88 cDNA is composed of 1682 bp and encodes a deduced polypeptide of 289 amino acids with 86% homology to MyD88 of Siniperca chuatsi.The deduced amino acid sequence of Og-MyD88 contains a typical death domain at the amino terminus and a conserved Toll/IL-1R(TIR) domain at the carboxyl terminus,as well as three highly conserved motifs(Box1,Box2 and Box3) within the C-terminal TIR domain.In healthy fish,Og-MyD88 was found to be strongly expressed in immune-related tissues,including the spleen,head kidney,kidney,liver,skin and intestine,with lower expression in heart,stomach,brain and muscle.Transcripts of Og-MyD88 were found to be markedly up-regulated in fish spleen after challenge with Singapore grouper iridovirus(SGIV),a highly lethal viral pathogen to grouper fish.Furthermore,the full length Og-MyD88 and its N-terminal death domain were capable of inducing NF-κB activity in HEK-293 cells.Overexpressed Og-MyD88 showed the ability to inhibit replication of SGIV in grouper spleen(GS) cells.These results suggest that Og-MyD88 is involved in the grouper immune response to invasion of viral pathogens and may share similar functions to those observed in higher vertebrates.展开更多
基金supported by the National Natural Science Foundation of China(31930115,32173007)China Agriculture Research System of MOF and MARA(CARS-47-G16)Basic and Applied Basic Research Foundation of Guangdong Province(2022A1515010595)。
文摘Iridovirus poses a substantial threat to global aquaculture due to its high mortality rate;however,the molecular mechanisms underpinning its pathogenesis are not well elucidated.Here,a multi-omics approach was applied to groupers infected with Singapore grouper iridovirus(SGIV),focusing on the roles of key metabolites.Results showed that SGIV induced obvious histopathological damage and changes in metabolic enzymes within the liver.Furthermore,SGIV significantly reduced the contents of lipid droplets,triglycerides,cholesterol,and lipoproteins.Metabolomic analysis indicated that the altered metabolites were enriched in 19 pathways,with a notable down-regulation of lipid metabolites such as glycerophosphates and alpha-linolenic acid(ALA),consistent with disturbed lipid homeostasis in the liver.Integration of transcriptomic and metabolomic data revealed that the top enriched pathways were related to cell growth and death and nucleotide,carbohydrate,amino acid,and lipid metabolism,supporting the conclusion that SGIV infection induced liver metabolic reprogramming.Further integrative transcriptomic and proteomic analysis indicated that SGIV infection activated crucial molecular events in a phagosome-immune depression-metabolism dysregulation-necrosis signaling cascade.Of note,integrative multi-omics analysis demonstrated the consumption of ALA and linoleic acid(LA)metabolites,and the accumulation of L-glutamic acid(GA),accompanied by alterations in immune,inflammation,and cell death-related genes.Further experimental data showed that ALA,but not GA,suppressed SGIV replication by activating antioxidant and anti-inflammatory responses in the host.Collectively,these findings provide a comprehensive resource for understanding host response dynamics during fish iridovirus infection and highlight the antiviral potential of ALA in the prevention and treatment of iridoviral diseases.
文摘Virus infection consists of entry, synthesis of macro- molecular components, virus assembly and release. Understanding of the mechanisms underlying each event is necessary for the intervention of virus infection in human healthcare and agriculture. Here we report the visualization of Singapore grouper iridovirus （SGIV） assembly in the medaka haploid embryonic stem （ES） cell line HX1. SGIV is a highly infectious DNA virus that causes a massive loss in marine aquaculture. Ectopic expression of VP88GFP, a fusion between green fluo- rescent protein and the envelope protein VP088, did not compromise the ES cell properties and susceptibility to SGIV infection. Although VP88GFP disperses evenly in the cytoplasm of non-infected cells, it undergoes aggregation and redistribution in SGIV-infected cells. Real-time visualization revealed multiple key stages of VP88GFP redistribution and the dynamics of viral assembly site （VAS）. Specifically, VP88GFP entry into and condensation in the VAS occurred within a 6-h duration, a similar duration was observed also for the release of VP88GFP-containing SGIV out of the cell, Taken together, VP088 is an excellent marker for visu- alizing the SGIV infection process. Our results provide new insight into macromolecular component recruit- ment and SGIV assembly.
基金supported by the National Basic Research Program of China (2012CB114402)the Knowledge Innovation Program of the Chinese Academy of Sciences (KZCX2-YW-BR-08 and KZCX2- EW-Q213)the National Natural Science Foundation of China (30725027 and 30930070)
文摘Myeloid differentiation factor 88(MyD88) is a universal adaptor protein involved in Toll-like receptors and in interleukin-1 receptor-induced nuclear factor-kappaB(NF-κB) activation.In this study,a new MyD88 gene(designated as Og-MyD88) was cloned from orange-spotted grouper,Epinephelus coioides,based on the expressed sequence tag(EST) obtained following Roche 454 GS-FLX sequencing.The full-length Og-MyD88 cDNA is composed of 1682 bp and encodes a deduced polypeptide of 289 amino acids with 86% homology to MyD88 of Siniperca chuatsi.The deduced amino acid sequence of Og-MyD88 contains a typical death domain at the amino terminus and a conserved Toll/IL-1R(TIR) domain at the carboxyl terminus,as well as three highly conserved motifs(Box1,Box2 and Box3) within the C-terminal TIR domain.In healthy fish,Og-MyD88 was found to be strongly expressed in immune-related tissues,including the spleen,head kidney,kidney,liver,skin and intestine,with lower expression in heart,stomach,brain and muscle.Transcripts of Og-MyD88 were found to be markedly up-regulated in fish spleen after challenge with Singapore grouper iridovirus(SGIV),a highly lethal viral pathogen to grouper fish.Furthermore,the full length Og-MyD88 and its N-terminal death domain were capable of inducing NF-κB activity in HEK-293 cells.Overexpressed Og-MyD88 showed the ability to inhibit replication of SGIV in grouper spleen(GS) cells.These results suggest that Og-MyD88 is involved in the grouper immune response to invasion of viral pathogens and may share similar functions to those observed in higher vertebrates.
