Host factors interacting with the pestivirus N erminal protease, Npro are components of the ribonucleoprotein complex

Jefferson, Matthew, Donaszi-Ivanov, Andras, Pollen, Sean, Dalmay, Tamas ORCID: https://orcid.org/0000-0003-1492-5429, Saalbach, Gerhard and Powell, Penny P. ORCID: https://orcid.org/0000-0002-5347-0490 (2014) Host factors interacting with the pestivirus N erminal protease, Npro are components of the ribonucleoprotein complex. Journal of Virology, 88 (18). pp. 10340-10353. ISSN 0022-538X

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Abstract

The viral N-terminal protease Npro of pestiviruses counteracts cellular antiviral defenses through inhibition of IRF3. Here we used mass spectrometry to identify a new role for Npro through its interaction with over 55 associated proteins, mainly ribosomal proteins and ribonucleoproteins, including RNA helicase A (DHX9), Y-box binding protein (YBX1), DDX3, DDX5, eIF3, IGF2BP1, multiple myeloma tumor protein 2, interleukin enhancer binding factor 3 (IEBP3), guanine nucleotide binding protein 3, and polyadenylate-binding protein 1 (PABP-1). These are components of the translation machinery, ribonucleoprotein particles (RNPs), and stress granules. Significantly, we found that stress granule formation was inhibited in MDBK cells infected with a noncytopathic bovine viral diarrhea virus (BVDV) strain, Kyle. However, ribonucleoproteins binding to Npro did not inhibit these proteins from aggregating into stress granules. Npro interacted with YBX1 though its TRASH domain, since the mutant C112R protein with an inactive TRASH domain no longer redistributed to stress granules. Interestingly, RNA helicase A and La autoantigen relocated from a nuclear location to form cytoplasmic granules with Npro. To address a proviral role for Npro in RNP granules, we investigated whether Npro affected RNA interference (RNAi), since interacting proteins are involved in RISC function during RNA silencing. Using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) silencing with small interfering RNAs (siRNAs) followed by Northern blotting of GAPDH, expression of Npro had no effect on RNAi silencing activity, contrasting with other viral suppressors of interferon. We propose that Npro is involved with virus RNA translation in the cytoplasm for virus particle production, and when translation is inhibited following stress, it redistributes to the replication complex.

Item Type:	Article
Additional Information:	Copyright © 2014 Jefferson et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 3.0 Unported license.
Faculty \ School:	Faculty of Medicine and Health Sciences > Norwich Medical School Faculty of Science > School of Biological Sciences Faculty of Science
UEA Research Groups:	Faculty of Science > Research Groups > Plant Sciences Faculty of Medicine and Health Sciences > Research Groups > Gastroenterology and Gut Biology Faculty of Medicine and Health Sciences > Research Centres > Metabolic Health
Depositing User:	Pure Connector
Date Deposited:	01 Aug 2014 14:22
Last Modified:	19 Oct 2023 01:20
URI:	https://ueaeprints.uea.ac.uk/id/eprint/49303
DOI:	10.1128/JVI.00984-14

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