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Virus Res ; 76(1): 17-29, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11376843

ABSTRACT

Previous results from our laboratory have identified a small (60 nt) RNA from the yeast S. cerevisiae that specifically inhibits internal ribosome entry site (IRES)-mediated translation programmed by poliovirus (PV) and hepatitis C virus (HCV) 5'-untranslated region (5'UTR). The yeast inhibitor RNA (called IRNA) was found to efficiently compete with viral 5'UTR for binding of several cellular polypeptides that presumably play important roles in IRES-mediated translation. One such IRNA (and 5'UTR)-binding protein has previously been identified as the La autoantigen. In this report, we have identified a 110-kDa IRNA-binding protein (which also interacts with viral 5'UTR) as nucleolin, a nucleolar RNA binding protein that was previously shown to translocate into the cytoplasm following infection of cells with poliovirus. We demonstrate that nucleolin (called C23) stimulates viral IRES-mediated translation both in vitro and in vivo. We also show that nucleolin mutants containing the carboxy-terminal RNA binding domains but lacking the amino terminal domain inhibit IRES-mediated translation in vitro. The translation inhibitory activity of these mutants correlates with their ability to bind the 5'UTR sequence. These results suggest a role of nucleolin/C23 in viral IRES-mediated translation.


Subject(s)
Gene Expression Regulation, Viral , Phosphoproteins/metabolism , Protein Biosynthesis , RNA-Binding Proteins/metabolism , Ribosomes/metabolism , 5' Untranslated Regions/genetics , 5' Untranslated Regions/metabolism , Genes, Dominant/genetics , HeLa Cells , Humans , Molecular Weight , Nuclear Proteins/chemistry , Nuclear Proteins/genetics , Nuclear Proteins/metabolism , Phosphoproteins/chemistry , Phosphoproteins/genetics , Poliovirus/genetics , Protein Binding , Protein Structure, Tertiary , RNA, Fungal/genetics , RNA, Fungal/metabolism , RNA-Binding Proteins/chemistry , RNA-Binding Proteins/genetics , Saccharomyces cerevisiae/genetics , Sequence Deletion , Viral Proteins/biosynthesis , Viral Proteins/genetics , Nucleolin
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