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High fidelity of yellow fever virus RNA polymerase

Konstantin V Pugachev1, Farshad Guirakhoo, Simeon W Ocran

  • 1Acambis, Inc., Cambridge, Massachusetts 02139, USA.

Journal of Virology
|December 25, 2003
PubMed

Insights

Researchers assessed the mutation rates of yellow fever virus-dengue virus vaccine candidates. The study found significant differences in error rates between in vitro transcription and viral genome replication, identifying key proteins for flavivirus assembly.

Area of Science:

  • Virology
  • Molecular Biology
  • Vaccine Development

Background:

  • Chimeric yellow fever virus-dengue virus (ChimeriVax-DEN) vaccine candidates were developed against dengue virus types 1-4.
  • Understanding viral genome replication fidelity is crucial for vaccine safety and efficacy.

Purpose of the Study:

  • To determine the nucleotide sequence error rates of SP6 RNA polymerase and yellow fever virus RNA polymerase in ChimeriVax-DEN vaccine candidates.
  • To identify potential protein regions involved in flavivirus assembly.

Main Methods:

  • Three consecutive plaque purifications of four ChimeriVax-DEN vaccine candidates.
  • Genome sequencing using a consensus approach after plaque purification and cell culture passages.
  • Analysis of nucleotide sequence error rates and mutation accumulation.

Main Results:

  • The nucleotide sequence error rate for SP6 RNA polymerase was high (1.34 x 10(-4) per copied nucleotide).
  • The error rate for yellow fever virus RNA polymerase in infected cells was low (1.9 x 10(-7) to 2.3 x 10(-7)).
  • Beneficial mutations clustered in the prM protein, E protein, and NS4B protein, suggesting their role in nucleocapsid-envelope interaction.

Conclusions:

  • Significant differences exist in the error rates of viral RNA polymerases during vaccine candidate replication.
  • The N-terminal prM, middle E protein, and NS4B protein are likely essential for flavivirus assembly.
  • These findings contribute to understanding flavivirus replication fidelity and assembly mechanisms.

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