Direct measurement of the poliovirus RNA polymerase error frequency in vitro
Open accessAbstract
The fidelity of RNA replication by the poliovirus-RNA-dependent RNA polymerase was examined by copying homopolymeric RNA templates in vitro. The poliovirus RNA polymerase was extensively purified and used to copy poly(A), poly(C), or poly(I) templates with equimolar concentrations of noncomplementary and complementary ribonucleotides. The error frequency was expressed as the amount of a noncomplementary nucleotide incorporated divided by the total amount of complementary and noncomplementary nucleotide incorporated. The polymerase error frequencies were very high and ranged from 7 x 10(-4) to 5.4 x 10(-3), depending on the specific reaction conditions. There were no significant differences among the error frequencies obtained with different noncomplementary nucleotide substrates on a given template or between the values determined on two different templates for a specific noncomplementary substrate. The activity of the polymerase on poly(U) and poly(G) was too low to measure error frequencies on these templates. A fivefold increase in the error frequency was observed when the reaction conditions were changed from 3.0 mM Mg2+ (pH 7.0) to 7.0 mM Mg2+ (pH 8.0). This increase in the error frequency correlates with an eightfold increase in the elongation rate that was observed under the same conditions in a previous study.
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The high mutation rate of poliovirus is in the order of 10−4 errors per nucleotide per replication cycle, due to the low fidelity of poliovirus RNA polymerase 3Dpol and the absence of a mismatch repair mechanism.
“The high mutation rate of PV is in the order of 10−4 errors per nt per replication cycle of the viral genome, due to the low fidelity of PV RNA polymerase 3D (3Dpol) and the absence of mismatch repair mechanism [15].”
From Recombination between Poliovirus and Coxsackie A Viruses of Species C: A Model of Viral Genetic Plasticity and Emergence (2011) · cites this paper as [15]
The rate of poliovirus evolution is extremely high and mainly due to the high frequency of nucleotide misincorporation in RNA synthesis.
“As mentioned above, the rate of PV evolution is extremely high and mainly due to the high frequency of nt misincorporation in RNA synthesis, [15].”
From Recombination between Poliovirus and Coxsackie A Viruses of Species C: A Model of Viral Genetic Plasticity and Emergence (2011) · cites this paper as [15]
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Lack of evidence for proofreading mechanisms associated with an RNA virus polymerase
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- Authors
- Ward, C. D., Stokes, M. A., & Flanegan, J. B.
- Affiliations
- University of Florida Health (US)
- Venue
- Journal of Virology
- Publisher
- American Society for Microbiology
- Published
- 1988-02
- Volume
- 62
- Issue
- 2
- Pages
- 558-562
- ISSN
- 0022-538X (ISSN-L)
- 1098-5514
- PMC
- PMC250568
- OpenAlex
- W1559971981
- Cited by
- 118
- References
- 35
- Citations by year
- 2025: 2 · 2023: 1 · 2022: 1 · 2021: 3 · 2020: 5 · 2019: 4 · 2018: 5 · 2017: 3 · 2016: 4 · 2015: 2 · 2014: 8 · 2013: 3 · 2012: 1
- FWCI
- 7.14
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Virology (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institute of Allergy and Infectious Diseases (AI15539, AI00523)
- Topics
- Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Viral Infections and Immunology Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Evolution and Genetic Dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Keywords
- error rate, magnesium content, RNA replication, elongation rate
- MeSH terms
- Nucleotidases, RNA Nucleotidyltransferases, RNA-Dependent RNA Polymerase, RNA, Viral, Substrate Specificity, Templates, Genetic, Single-Strand Specific DNA and RNA Endonucleases, Poliovirus
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