The Npro product of classical swine fever virus interacts with IκBα, the NF-κB inhibitor

Abstract

Classical swine fever virus(CSFV) belongs to the genusPestivirusand is the causative agent of classical swine fever, a haemorrhagic disease of pigs. The virus replicates in host cells without activating interferon (IFN) production and has been reported to be an antagonist of double-stranded RNA-induced apoptosis. The N-terminal protease (Npro) of CSFV is responsible for this evasion of the host innate immune response. In order to identify cellular proteins that interact with the Nproproduct of CSFV, a yeast two-hybrid screen of a human library was carried out, which identified IκBα, the inhibitor of NF-κB, a transcription factor involved in the control of apoptosis, the immune response and IFN production. The Npro–IκBαinteraction was confirmed using yeast two-hybrid analysis and additional co-precipitation assays. It was also shown that Nprolocalizes to both the cytoplasmic and nuclear compartments in stably transfected cells and in CSFV-infected cells. Following stimulation by tumour necrosis factor alpha, PK-15 cell lines expressing Nproexhibited transient nuclear accumulation of pIκBα, but no effect of CSFV infection on IκBαlocalization or NF-κB p65 activation was observed.