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Identifying Caspases and their Motifs that Cleave Proteins During Influenza A Virus Infection
Published on: July 21, 2022
Molecular characterization of caspase members and expression response to Nervous Necrosis Virus outbreak in Pacific
Ming-Guang Mao1, Jie-Lan Jiang2, Zhi-Qiang Jiang2
1State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; Key Laboratory of Mariculture and Stock Enhancement in North China's Sea, Ministry of Agriculture, Dalian Ocean University, Dalian 262306, China.
Abstract:
Multiple functions of caspases include normal cell turnover, proper development and function of the immune system, hormone-dependent atrophy, embryonic development and chemical-induced cell injury. During artificial propagation of Pacific cod, Gadus macrocephalus, high mortality occurred during early development stages. Here, we performed various analyses on the cDNA and protein sequences of six different G. macrocephalus caspases namely GmCasp3, 6, 7, 8, 9 and 10, and tried to investigate the contributions of caspase family to the development and Nervous Necrosis Virus (NNV) resistance. Sequence analysis of GmCaspase proteins showed that each caspase shared conserved domains like "HG", "QACXG (X for R, G or Q)" and "GSWF" except GmCasp10. Sequence alignment and phylogenetic tree showed that GmCasp8 and GmCasp10 were quite different from those of other fishes. 3-D models indicated that structure of GmCasp3 is very conservative, but GmCasp6, 7, 8, 9 and 10 are less conservative. Tissue distribution analysis showed that six Gmcaspases mRNA transcripts were detected in tissues of intestine, gill, thymus, head-kidney and spleen with different abundance, but Gmcasp7 were not detected in the brain. GmCasp3 transcript was kept at very low level in the early development stages, while the expression levels of GmCasp6, 7, 8, 10 were different at various development stages. GmCasp8 level seemed to be much higher than other caspases in the heads of 65dph and 75dph juveniles. To understand the role of caspases during NNV outbreak, modulation in expression of each Gmcaspases were investigated. The results showed that GmCasp3 transcript level increased significantly when NNV broke out, while GmCasp7, 8, 9 and 10 in cod heads decreased obviously at 69dph and 77dph. The results suggest that caspases in Pacific cod should be diverse in their structure and function, and their unique features and response to NNV outbreak add more evidences for the specificity of immune system in Pacific cod.
Insights
Pacific cod caspases (GmCaspases) show diverse structures and functions, with GmCasp3 levels rising during Nervous Necrosis Virus outbreaks, while others decrease, highlighting immune system specificity.
Area of Science:
- Molecular Biology
- Immunology
- Aquaculture
Background:
- Caspases are crucial enzymes involved in various cellular processes, including immune responses and development.
- Pacific cod (Gadus macrocephalus) aquaculture faces challenges with high mortality during early development stages.
- Understanding caspase function is vital for improving fish health and disease resistance, particularly against Nervous Necrosis Virus (NNV).
Purpose of the Study:
- To analyze the cDNA and protein sequences of six Pacific cod caspases (GmCasp3, 6, 7, 8, 9, and 10).
- To investigate the structural diversity, tissue distribution, developmental expression, and response to NNV infection of these caspases.
- To elucidate the role of caspases in Pacific cod development and NNV resistance.
Main Methods:
- Sequence analysis of GmCaspase cDNA and protein sequences.
- Phylogenetic analysis and 3-D modeling of caspase structures.
- Tissue distribution analysis of Gmcaspase mRNA transcripts via quantitative PCR.
- Expression profiling of Gmcaspases during different developmental stages and NNV infection.
Main Results:
- GmCaspase proteins exhibited conserved domains, except GmCasp10. GmCasp8 and GmCasp10 showed significant divergence from other fish species.
- Structural analysis revealed GmCasp3 as highly conserved, while others were less so. Gmcaspase transcripts were detected in various tissues, with Gmcasp7 absent in the brain.
- GmCasp3 expression was low during early development but increased significantly during NNV outbreaks. GmCasp7, 8, 9, and 10 levels decreased in NNV-infected cod heads.
Conclusions:
- Pacific cod caspases display structural and functional diversity, contributing to the specificity of their immune system.
- The differential expression patterns of Gmcaspases during development and NNV infection suggest distinct roles in cellular processes and disease response.
- These findings provide insights into caspase functions in fish, aiding in the development of strategies to enhance aquaculture resilience.
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