Comparative Proteomic Analysis of Lysine Acetylation in Fish CIK Cells Infected with Aquareovirus

Abstract

Grass carp (Ctenopharyngodon idellus) is an important worldwide commercial freshwater culture species. However, grass carp reovirus (GCRV) causes serious hemorrhagic disease in fingerlings and yearlings of fishes. To understand the molecular pathogenesis of host cells during GCRV infection, intensive proteomic quantification analysis of lysine acetylation in Ctenopharyngodon idella kidney (CIK) cells was performed. Using dimethylation labeling-based quantitative proteomics, 832 acetylated proteins with 1391 lysine acetylation sites were identified in response to GCRV infection, among which 792 proteins with 1323 sites were quantifiable. Bioinformatics analysis showed that differentially expressed lysine acetylated proteins are involved in diverse cellular processes and associated with multifarious functions, suggesting that extensive intracellular activities were changed upon viral infection. In addition, extensive alterations on host–protein interactions at the lysine acetylation level were also detected. Further biological experiments showed that the histone deacetylases (HDAC) inhibitor suberoylanilide hydroxamic acid (SAHA) could significantly suppress the GCRV replication. To our knowledge, this is the first to reveal the proteome-wide changes in host cell acetylome with aquatic virus infection. The results provided in this study laid a basis for further understanding the host response to aquareovirus infection in the post-translational modification aspect by regulating cell lysine acetylation conducive to viral replication.

Orchestration of protein acetylation as a toggle for cellular defense and virus replication

“…The presence of PTMs on viral proteins adds another layer of complexity to the role of acetylation during infection. Although recent work in insect and fish species revealed viral acetylomes as critical contributors to infection 67 , 68 , acetylation of viral proteins during human pathogenic infections remains understudied. Most of our understanding of viral protein acetylation comes from work on individual proteins, such as influenza A NP, HIV Tat, and papillomavirus E2 6 , 8 , 10 .…”

Integrative Proteome and Acetylome Analyses of Murine Responses to Cryptococcus neoformans Infection

“…Brain and lung tissue acetylome measurements were performed in biological triplicates as described elsewhere . A proteome analysis was performed first, wherein the proteins were digested with trypsin as described elsewhere (Guo et al, 2017;. Briefly, tissue protein samples were collected and reduced with 5 mM DTT for 30 min at 56 • C and alkylated using 11 mM iodoacetimide for 15 min at room temperature in darkness, then diluted with 0.1 M TEAB to normalize protein concentration across all samples, and then digested with trypsin (Promega, Madison, CT, United States) at 37 • C overnight.…”

Protein–protein interaction network: an emerging tool for understanding fish disease in aquaculture

“…Grass carp reovirus (GCRV) causes severe haemorrhagic disease in grass carp Ctenopharyngodon idellusa common freshwater aquaculture species. Following GCRV infection, 832 acetylated proteins with 1391 lysine acetylation sites were identified by proteomic quantification analysis in kidney cells (Guo et al 2017a). PPI network of 187 differentially expressed acetylated proteins was constructed, and extensive interruption on host-protein interactions was detected at the acetylation level in host cells upon viral infection.…”