High Fidelity of Yellow Fever Virus RNA Polymerase

Konstantin V. Pugachev, Farshad Guirakhoo, Simeon W. Ocran, Fred Mitchell, Megan Parsons, Caroline Penal, Soheila Girakhoo, Svetlana O. Pougatcheva, Juan Arroyo, Dennis W. Trent, Thomas P. Monath

Research output: Contribution to journalArticle

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Abstract

Three consecutive plaque purifications of four chimeric yellow fever virus-dengue virus (ChimeriVax-DEN) vaccine candidates against dengue virus types 1 to 4 were performed. The genome of each candidate was sequenced by the consensus approach after plaque purification and additional passages in cell culture. Our data suggest that the nucleotide sequence error rate for SP6 RNA polymerase used in the in vitro transcription step to initiate virus replication was as high as 1.34 × 10-4 per copied nucleotide and that the error rate of the yellow fever virus RNA polymerase employed by the chimeras for genome replication in infected cells was as low as 1.9 × 10-7 to 2.3 × 10-7. Clustering of beneficial mutations that accumulated after multiple virus passages suggests that the N-terminal part of the prM protein, a specific site in the middle of the E protein, and the NS4B protein may be essential for nucleocapsid-envelope interaction during flavivirus assembly.

Original languageEnglish (US)
Pages (from-to)1032-1038
Number of pages7
JournalJournal of Virology
Volume78
Issue number2
DOIs
StatePublished - Jan 2004

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Yellow fever virus
DNA-Directed RNA Polymerases
DNA-directed RNA polymerase
Dengue virus
Dengue Virus
Genome
Flavivirus
Nucleocapsid
nucleocapsid
Proteins
genome
proteins
chimerism
Virus Replication
virus replication
Cluster Analysis
cell culture
Vaccines
Nucleotides
transcription (genetics)

ASJC Scopus subject areas

  • Immunology

Cite this

Pugachev, K. V., Guirakhoo, F., Ocran, S. W., Mitchell, F., Parsons, M., Penal, C., ... Monath, T. P. (2004). High Fidelity of Yellow Fever Virus RNA Polymerase. Journal of Virology, 78(2), 1032-1038. https://doi.org/10.1128/JVI.78.2.1032-1038.2004

High Fidelity of Yellow Fever Virus RNA Polymerase. / Pugachev, Konstantin V.; Guirakhoo, Farshad; Ocran, Simeon W.; Mitchell, Fred; Parsons, Megan; Penal, Caroline; Girakhoo, Soheila; Pougatcheva, Svetlana O.; Arroyo, Juan; Trent, Dennis W.; Monath, Thomas P.

In: Journal of Virology, Vol. 78, No. 2, 01.2004, p. 1032-1038.

Research output: Contribution to journalArticle

Pugachev, KV, Guirakhoo, F, Ocran, SW, Mitchell, F, Parsons, M, Penal, C, Girakhoo, S, Pougatcheva, SO, Arroyo, J, Trent, DW & Monath, TP 2004, 'High Fidelity of Yellow Fever Virus RNA Polymerase', Journal of Virology, vol. 78, no. 2, pp. 1032-1038. https://doi.org/10.1128/JVI.78.2.1032-1038.2004
Pugachev KV, Guirakhoo F, Ocran SW, Mitchell F, Parsons M, Penal C et al. High Fidelity of Yellow Fever Virus RNA Polymerase. Journal of Virology. 2004 Jan;78(2):1032-1038. https://doi.org/10.1128/JVI.78.2.1032-1038.2004
Pugachev, Konstantin V. ; Guirakhoo, Farshad ; Ocran, Simeon W. ; Mitchell, Fred ; Parsons, Megan ; Penal, Caroline ; Girakhoo, Soheila ; Pougatcheva, Svetlana O. ; Arroyo, Juan ; Trent, Dennis W. ; Monath, Thomas P. / High Fidelity of Yellow Fever Virus RNA Polymerase. In: Journal of Virology. 2004 ; Vol. 78, No. 2. pp. 1032-1038.
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