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Updated: Jul 19, 2025
18:10
Isolation of Fidelity Variants of RNA Viruses and Characterization of Virus Mutation Frequency
Published on: June 16, 2011
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Single-cell mutation rate of turnip crinkle virus (-)-strand replication intermediates
Camila Perdoncini Carvalho1,2, Junping Han1, Khwannarin Khemsom1
1Department of Plant Pathology, The Ohio State University, Wooster, Ohio, United States of America.
Plos Pathogens
|August 14, 2023
Summary
Turnip crinkle virus (TCV) replication error rates were precisely measured using a novel method. This study reveals TCV
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- Positive-sense single-stranded RNA viruses exhibit high replication error rates, driving viral evolution and adaptation.
- Accurately quantifying these error rates in (+) RNA viruses is challenging due to natural selection and experimental artifacts.
- Turnip crinkle virus (TCV) is a model (+) RNA plant virus where precise error rate determination is needed.
Purpose of the Study:
- To develop and apply a novel method for accurately measuring the replication error rate of TCV.
- To investigate the fidelity of TCV RNA-dependent RNA polymerase and replication mechanisms.
- To understand the role of replication errors in viral genome diversification and adaptation.
Main Methods:
- Developed a new approach to profile errors specifically in negative-sense (-) RNA replication intermediates of TCV.
- Utilized singly infected cells to minimize confounding factors.
- Implemented controls and safeguards to account for experimental process errors and ensure accurate error rate estimation.
Main Results:
- Estimated the TCV substitution error rate at 8.47 x 10^-5 per nucleotide site per cell infection.
- Error distribution patterns indicated that most TCV (-) strands are products of a single replication cycle.
- Identified potential mechanisms, including polymerase fidelity and re-replication, influencing error frequencies.
Conclusions:
- The study provides a robust method for quantifying (+) RNA virus replication error rates.
- Findings support a 'stamping machine' mode of replication for TCV and potentially other (+) RNA viruses.
- Accurate error rate data is crucial for understanding viral evolution, adaptation, and developing antiviral strategies.
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