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The N^pro product of classical swine fever virus interacts with IB, the NF-B inhibitor

Virginie Doceul^1,

Penny P. Powell^1,

¹ BBSRC Institute for Animal Health, Pirbright Laboratory, Ash Road, Pirbright, Surrey GU24 0NF, UK
² Veterinary Laboratories Agency, New Haw, Addlestone, Surrey KT15 3NB, UK

Correspondence
Julian Seago
julian.seago{at}bbsrc.ac.uk

Classical swine fever virus (CSFV) belongs to the genus Pestivirus and 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 (N^pro) of CSFV is responsible for this evasion of the host innate immune response. In order to identify cellular proteins that interact with the N^pro product of CSFV, a yeast two-hybrid screen of a human library was carried out, which identified IB, the inhibitor of NF-B, a transcription factor involved in the control of apoptosis, the immune response and IFN production. The N^pro–IB interaction was confirmed using yeast two-hybrid analysis and additional co-precipitation assays. It was also shown that N^pro localizes 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 N^pro exhibited transient nuclear accumulation of pIB, but no effect of CSFV infection on IB localization or NF-B p65 activation was observed.

Present address: Department of Virology, Faculty of Medicine, Imperial College London, St Mary's Campus, Norfolk Place, London W2 1PG, UK.

Present address: University of East Anglia, Norwich NR4 7TJ, UK.

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