RNA-dependent RNA polymerase speed and fidelity are not the only determinants of the mechanism or efficiency of recombination
Public Deposited- Creator
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Kim, H.
- School of Medicine, Department of Microbiology and Immunology
- Other Affiliation: The Pennsylvania State University
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Ellis, V.D., III
- Other Affiliation: The Pennsylvania State University
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Woodman, A.
- Other Affiliation: The Pennsylvania State University
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Zhao, Y.
- Other Affiliation: The Pennsylvania State University
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Arnold, J.J.
- School of Medicine, Department of Microbiology and Immunology
- Other Affiliation: The Pennsylvania State University
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Cameron, C.E.
- School of Medicine, Department of Microbiology and Immunology
- Other Affiliation: The Pennsylvania State University
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Kim, H.
- Abstract
- Using the RNA-dependent RNA polymerase (RdRp) from poliovirus (PV) as our model system, we have shown that Lys-359 in motif-D functions as a general acid in the mechanism of nucleotidyl transfer. A K359H (KH) RdRp derivative is slow and faithful relative to wild-type enzyme. In the context of the KH virus, RdRp-coding sequence evolves, selecting for the following substitutions: I331F (IF, motif-C) and P356S (PS, motif-D). We have evaluated IF-KH, PS-KH, and IF-PS-KH viruses and enzymes. The speed and fidelity of each double mutant are equivalent. Each exhibits a unique recombination phenotype, with IF-KH being competent for copy-choice recombination and PS-KH being competent for forced-copy-choice recombination. Although the IF-PS-KH RdRp exhibits biochemical properties within twofold of wild type, the virus is impaired substantially for recombination in cells. We conclude that there are biochemical properties of the RdRp in addition to speed and fidelity that determine the mechanism and efficiency of recombination. The interwoven nature of speed, fidelity, the undefined property suggested here, and recombination makes it impossible to attribute a single property of the RdRp to fitness. However, the derivatives described here may permit elucidation of the importance of recombination on the fitness of the viral population in a background of constant polymerase speed and fidelity.
- Date of publication
- 2019
- DOI
- Identifier
- https://dx.doi.org/10.3390/genes10120968
- PMID 31775299
- Resource type
- Article
- Rights statement
- In Copyright
- Journal title
- Genes
- Journal volume
- 10
- Journal issue
- 12
- Language
- English
- ISSN
- 2073-4425
- Publisher
- MDPI AG
Relations
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