title: High fidelity of yellow fever virus RNA polymerase.
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 x 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 x 10(-7) to 2.3 x 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.
authors.affiliation: Acambis, Inc., Cambridge, Massachusetts 02139, USA.
authors.forename: Konstantin V
authors.initials: KV
authors.lastname: Pugachev
authors.forename: Farshad
authors.initials: F
authors.lastname: Guirakhoo
authors.forename: Simeon W
authors.initials: SW
authors.lastname: Ocran
authors.forename: Fred
authors.initials: F
authors.lastname: Mitchell
authors.forename: Megan
authors.initials: M
authors.lastname: Parsons
authors.forename: Caroline
authors.initials: C
authors.lastname: Penal
authors.forename: Soheila
authors.initials: S
authors.lastname: Girakhoo
authors.forename: Svetlana O
authors.initials: SO
authors.lastname: Pougatcheva
authors.forename: Juan
authors.initials: J
authors.lastname: Arroyo
authors.forename: Dennis W
authors.initials: DW
authors.lastname: Trent
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Monath
mesh_terms: D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D003715:Dengue / Q000517:prevention & control; D003716:Dengue Virus / Q000235:genetics; D017422:Sequence Analysis, DNA; D012692:Serial Passage; D014158:Transcription, Genetic; D014614:Vaccines, Synthetic; D010948:Viral Plaque Assay; D014765:Viral Vaccines; D019065:Virus Assembly; D014779:Virus Replication; D015005:Yellow fever virus / Q000201:enzymology / Q000235:genetics
chemical_list: D014614:Vaccines, Synthetic; D014765:Viral Vaccines; D012321:DNA-Directed RNA Polymerases