Home Acta Virologica 2005 Acta Virologica Vol.49, p.83-90, 2005

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Quarterly,
Founded: 1957
ISSN 0001-723X
E-ISSN 1336-2305

Published in English

Impact Factor = 1.82

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Acta Virologica Vol.49, p.83-90, 2005

Title: TWO CIS-ACTING ELEMENTS IN NEGATIVE RNA STRAND OF HEPATITIS C VIRUS INVOLVED IN SYNTHESIS OF POSITIVE RNA STRAND IN VITRO
Author: L. YE, K.A. TIMANI, L. YE, L. KONG, X. YANG, Q. LIAO, J. WU

Abstract: Sequences at the 3'-ends of both positive and negative strands of Hepatitis C virus (HCV) RNA harbor cis-acting elements required for RNA replication. However, little is known about the properties of the negative RNA strand as a template for the synthesis of positive RNA strand. In this study, a purified recombinant HCV RNA-dependent RNA polymerase (RdRp) was used to investigate the synthesis of positive RNA strand using the 3'-terminal region of negative RNA strand ((–)3'T RNA) as template. A mutagenesis analysis was performed to evaluate the role of the 3'-proximal stem-loop and the first 3'-cytidylate (3'C) of the negative RNA strand in the synthesis of the positive RNA strand. A negative RNA strand of wild type (wt) HCV as template was able to direct the synthesis of a full-length positive RNA strand. Deletion of the 3'-proximal stem-loop resulted in an approximately 90% decrease in RNA synthesis. Disruption of the 3'-proximal stem-loop structure by nucleotide substitutions led to a 70–80% decrease in RNA synthesis. However, the restoration of the stem-loop by compensatory mutations in the stem region restored also the RNA synthesis. Likewise, the deletion or substitution of the first 3'C by guanylate (G) led to a 90% decrease in the RNA synthesis; while the substitution by adenylate (A) or uridylate (U) resulted in a 60–80% decrease in the RNA synthesis only. These findings demonstrate that the 3'-proximal stem-loop and the first 3'C of the negative RNA strand of HCV are two cis-acting elements involved in the synthesis of the positive RNA strand.

Keywords: Hepatitis C virus; RNA; positive RNA strand; negative RNA strand; RNA-dependent RNA polymerase; RNA synthesis; cis-acting element
Year: 2005, Volume: 49, Issue: Page From: 83, Page To: 90



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