HuR binding to AU-rich elements present in the 3′ untranslated region of Classical swine fever virus

Abstract

Background: Classical swine fever virus (CSFV) is the member of the genus Pestivirus under the family Flaviviridae. The 5′ untranslated region (UTR) of CSFV contains the IRES, which is a highly structured element that recruits the translation machinery. The 3′ UTR is usually the recognition site of the viral replicase to initiate minus-strand RNA synthesis. Adenosine-uridine rich elements (ARE) are instability determinants present in the 3′ UTR of short-lived mRNAs. However, the presence of AREs in the 3′ UTR of CSFV conserved in all known strains has never been reported. This study inspects a possible role of the ARE in the 3′ UTR of CSFV. Results: Using RNA pull-down and LC/MS/MS assays, this study identified at least 32 possible host factors derived from the cytoplasmic extracts of PK-15 cells that bind to the CSFV 3′ UTR, one of which is HuR. HuR is known to bind the AREs and protect the mRNA from degradation. Using recombinant GST-HuR, this study demonstrates that HuR binds to the ARE present in the 3′ UTR of CSFV in vitro and that the binding ability is conserved in strains irrespective of virulence. Conclusions: This study identified one of the CSFV 3′ UTR binding proteins HuR is specifically binding to in the ARE region. © 2011 Nadar et al; licensee BioMed Central Ltd.