Unwinding of nucleic acids by HCV NS3 helicase is sensitive to the structure of the duplex

Alan J. Tackett, Lai Wei, Craig Eugene Cameron, Kevin D. Raney

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

Hepatitis C virus (HCV) helicase, non-structural protein 3 (NS3), is proposed to aid in HCV genome replication and is considered a target for inhibition of HCV. In order to investigate the substrate requirements for nucleic acid unwinding by NS3, substrates were prepared by annealing a 30mer oligonucleotide to a 15mer. The resulting 15 bp duplex contained a single-stranded DNA overhang of 15 nt referred to as the bound strand. Other substrates were prepared in which the 15mer DNA was replaced by a strand of peptide nucleic acid (PNA). The PNA-DNA substrate was unwound by NS3, but the observed rate of strand separation was at least 25-fold slower than for the equivalent DNA-DNA substrate. Binding of NS3 to the PNA-DNA substrate was similar to the DNA-DNA substrate, due to the fact that NS3 initially binds to the single-stranded overhang, which was identical in each substrate. A PNA-RNA substrate was not unwound by NS3 under similar conditions. In contrast, morpholino-DNA and phosphorothioate-DNA substrates were utilized as efficiently by NS3 as DNA-DNA substrates. These results indicate that the PNA-DNA and PNA-RNA heteroduplexes adopt structures that are unfavorable for unwinding by NS3, suggesting that the unwinding activity of NS3 is sensitive to the structure of the duplex.

Original languageEnglish (US)
Pages (from-to)565-572
Number of pages8
JournalNucleic Acids Research
Volume29
Issue number2
StatePublished - Jan 15 2001

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Viral Structural Proteins
Hepacivirus
Nucleic Acids
Peptide Nucleic Acids
DNA
Proteins
Nucleic Acid Heteroduplexes
RNA
Morpholinos
Single-Stranded DNA
Virus Replication
Oligonucleotides

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

Tackett, A. J., Wei, L., Cameron, C. E., & Raney, K. D. (2001). Unwinding of nucleic acids by HCV NS3 helicase is sensitive to the structure of the duplex. Nucleic Acids Research, 29(2), 565-572.
Tackett, Alan J. ; Wei, Lai ; Cameron, Craig Eugene ; Raney, Kevin D. / Unwinding of nucleic acids by HCV NS3 helicase is sensitive to the structure of the duplex. In: Nucleic Acids Research. 2001 ; Vol. 29, No. 2. pp. 565-572.
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Tackett, AJ, Wei, L, Cameron, CE & Raney, KD 2001, 'Unwinding of nucleic acids by HCV NS3 helicase is sensitive to the structure of the duplex', Nucleic Acids Research, vol. 29, no. 2, pp. 565-572.

Unwinding of nucleic acids by HCV NS3 helicase is sensitive to the structure of the duplex. / Tackett, Alan J.; Wei, Lai; Cameron, Craig Eugene; Raney, Kevin D.

In: Nucleic Acids Research, Vol. 29, No. 2, 15.01.2001, p. 565-572.

Research output: Contribution to journalArticle

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