Differential Ubiquitination of Host Proteins in Response to IPNV and ISAV viruses in Atlantic salmon

Abstract

Viral diseases remain a major barrier to the sustainable production of farmed fish, primarily attributable to the absence of effective prevention and treatment options. Understanding host-pathogen interactions can guide the development of vaccines, antiviral therapies, or gene editing strategies. Ubiquitination is a key cell signalling molecule, known to regulate many aspects of immune functions but currently understudied in fish. This study leverages ubiquitin-enriched mass spectroscopy complemented with RNA sequencing to characterize the role of ubiquitination in response to infection. A challenge experiment was conducted by infecting Atlantic salmon head kidney (SHK-1) cells with Infectious salmon anaemia virus (ISAV) and Infectious pancreatic necrosis virus (IPNV). At 24- and 48 hours post-infection dramatic changes were observed in the global ubiquitination state of host proteins. Many post-translational modifying proteins increased in abundance upon ISAV infection, whilst IPNV infection resulted in a reduction in abundance of many of these proteins. Transcriptomics showed a delay in the activation of the antiviral response to ISAV infection, with major upregulation of genes associated with immune pathways only at 48h. On the contrary, IPNV infection resulted in upregulation of classic innate immune response genes at both timepoints. Clear activation of Rig-like receptor pathways is demonstrated in both infections, in addition to upregulation of both conserved and novel antiviral TRIM E3 ubiquitin ligate genes. Network analysis identified clusters of immune genes and putatively regulatory proteins showing differential ubiquitination upon viral infection. This study highlights the importance of post-translational control of the host innate immune response to viruses in Atlantic salmon. Clear differences in ubiquitination between two viruses indicate either virus-specific post-translational regulation or viral antagonism of the immune response. Additionally, the ubiquitination of various proteins was linked to the regulation of innate immune pathways, suggesting a direct role of ubiquitination in the regulation of antiviral responses. AUTHOR SUMMARYUbiquitination is a main cellular regulatory mechanism in all animal species, controlling the activity of many proteins within the cell. The role of ubiquitination in the regulation of immunity is well-established, and several direct interactions with viruses have been described, either as part of a strategy of the host to fight the infection or as a viral mechanism to evade the host immune response. In aquaculture, viral diseases currently represent the most important threat to the sustainability of the industry. However, very little is known about the interplay between ubiquitination and viral pathogens in fish species. In our study, we have assessed the ubiquitination response of Atlantic salmon to two production-relevant viruses, Infectious salmon anaemia virus (ISAV) and Infectious pancreatic necrosis virus (IPNV). We found remarkable differences in the ubiquitination profiles between the two infections, suggesting a key role of ubiquitination on the early immune response. We also discovered an association between the ubiquitination of certain regulatory proteins and the activation of antiviral pathways. This information could help develop new strategies to tackle viral diseases in aquaculture, such as the development of more effective vaccines or antiviral therapies, or inform gene editing efforts to generate disease resistant fish.