African Swine Fever Virus F317L Protein Inhibits NF-κB Activation To Evade Host Immune Response and Promote Viral Replication.

Abstract

ABSTRACTAfrican swine fever (ASF) is a highly contagious and deadly viral disease affecting pigs, with up to a 100% case fatality rate. The causative agent, African swine fever virus (ASFV), is a large multienveloped DNA virus which is the sole member of the family Asfarviridae. The double-stranded DNA genome of ASFV encodes more than 150 proteins; the functions of more than half of these viral proteins remain unknown. In this study, we determined that the uncharacterized protein F317L of ASFV had an antagonistic function against host innate immune response. F317L impaired NF-κB pathway activation by disruption of NF-κB activity. F317L interacted with IκB kinase β (IKKβ) and suppressed its phosphorylation, which subsequently reduced phosphorylation and ubiquitination of IκBα and enhanced IκBα stabilization. The accumulation of IκBα then blocked NF-κB activation and inhibited its nuclear translocation, resulting in decreased expression of various proinflammatory cytokines. As expected, overexpression of F317L promoted ASFV replication, and knockdown of F317L expression suppressed ASFV replication. This also indicated the crucial role of NF-κB pathway signaling in suppression of ASFV replication. Truncation mutation analysis indicated that the region spanning amino acids 109 to 208 of F317L was critical for inhibition of NF-κB activity. This is the first report about the function of F317L protein of ASFV, which provides insights for investigation of ASFV immune evasion mechanisms and development of ASFV live-attenuated vaccine.IMPORTANCE African swine fever (ASF) is one of the most important pig diseases, causing a high case fatality rate and trade restrictions upon reported outbreaks. The limited understanding of the functions of the proteins of the causative agent, African swine fever virus (ASFV), has become a primary barrier to developing available commercial ASFV vaccines. ASFV infection causes severe immunosuppression. However, the mechanisms are still poorly understood. Identification of the viral factors responsible for causing immunosuppression will provide targets for developing ASFV live-attenuated vaccine through deletion of these viral factors. In this study, we determined that the uncharacterized protein F317L of ASFV had an antagonistic function against host innate immune response. Knockdown of F317L expression clearly inhibited ASFV replication. This is the first report about the function of F317L protein of ASFV, which provides new data to understand how ASFV inhibits host innate immune response and provides insights for developing ASFV live-attenuated vaccine.