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CUSTOMERS Acta Virologica 2012 Acta Virologica Vol.56, No.3, p.217-226, 2012
CUSTOMERS Acta Virologica 2012 Acta Virologica Vol.56, No.3, p.217-226, 2012
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Acta Virologica Vol.56, No.3, p.217-226, 2012 |
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| Title: The adenine-rich tract in the 5ʹ-end of the hepatitis C virus ORF encodes a peptide regulating the binding of the C protein to RNA | ||
| Author: S. CHINNASWAMY, W. B. LOTT | ||
| Abstract: Hepatitis C virus (HCV) core (C) protein is thought to bind to viral RNA before it undergoes oligomerization leading to RNA encapsidation. Details of these events are so far unknown. The 5ʹ-terminal C protein coding sequence that includes an adenine (A)-rich tract is a part of an internal ribosome entry site (IRES). This nucleotide sequence but not the corresponding protein sequence is needed for proper initiation of translation of viral RNA by an IRES-dependent mechanism. In this study, we examined the importance of this sequence for the ability of the C protein to bind to viral RNA. Serially truncated C proteins with deletions from 10 up to 45 N-terminal amino acids were expressed in Escherichia coli, purified and tested for binding to viral RNA by a gel shift assay. The results showed that truncation of the C protein from its N-terminus by more than 10 amino acids abolished almost completely its expression in E. coli. The latter could be restored by adding a tag to the N-terminus of the protein. The tagged proteins truncated by 15 or more amino acids showed an anomalous migration in SDS-PAGE. Truncation by more than 20 amino acids resulted in a complete loss of ability of tagged C protein to bind to viral RNA. These results provide clues to the early events in the C protein - RNA interactions leading to C protein oligomerization, RNA encapsidation and virion assembly. |
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| Keywords: hepatitis C virus; core protein; RNA binding | ||
| Year: 2012, Volume: 56, Issue: 3 | Page From: 217, Page To: 226 | |
| doi:10.4149/av_2012_03_217 |
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