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Title: Direct measurement of the poliovirus RNA polymerase error frequency in vitro

Abstract

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, depending on the specific reaction conditions. 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 Mg{sup 2+} (pH 7.0) to 7.0 mM Mg{sup 2+} (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.

Authors:
; ;  [1]
  1. Univ. of Florida College of Medicine, Gainesville (USA)
Publication Date:
OSTI Identifier:
6525520
Resource Type:
Journal Article
Journal Name:
Journal of Virology; (USA)
Additional Journal Information:
Journal Volume: 62:2; Journal ID: ISSN 0022-538X
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; NUCLEOTIDES; AUTORADIOGRAPHY; POLIO VIRUS; NUCLEIC ACID REPLICATION; RNA POLYMERASES; PURIFICATION; BIOCHEMISTRY; ERRORS; PHOSPHORUS 32; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; CHEMISTRY; DAYS LIVING RADIOISOTOPES; ENZYMES; ISOTOPES; LIGHT NUCLEI; MICROORGANISMS; NUCLEI; NUCLEOTIDYLTRANSFERASES; ODD-ODD NUCLEI; ORGANIC COMPOUNDS; PARASITES; PHOSPHORUS ISOTOPES; PHOSPHORUS-GROUP TRANSFERASES; POLYMERASES; RADIOISOTOPES; TRANSFERASES; VIRUSES; 550701* - Microbiology- Tracer Techniques

Citation Formats

Ward, C D, Stokes, M A.M., and Flanegan, J B. Direct measurement of the poliovirus RNA polymerase error frequency in vitro. United States: N. p., 1988. Web.
Ward, C D, Stokes, M A.M., & Flanegan, J B. Direct measurement of the poliovirus RNA polymerase error frequency in vitro. United States.
Ward, C D, Stokes, M A.M., and Flanegan, J B. 1988. "Direct measurement of the poliovirus RNA polymerase error frequency in vitro". United States.
@article{osti_6525520,
title = {Direct measurement of the poliovirus RNA polymerase error frequency in vitro},
author = {Ward, C D and Stokes, M A.M. and Flanegan, J B},
abstractNote = {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, depending on the specific reaction conditions. 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 Mg{sup 2+} (pH 7.0) to 7.0 mM Mg{sup 2+} (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.},
doi = {},
url = {https://www.osti.gov/biblio/6525520}, journal = {Journal of Virology; (USA)},
issn = {0022-538X},
number = ,
volume = 62:2,
place = {United States},
year = {Mon Feb 01 00:00:00 EST 1988},
month = {Mon Feb 01 00:00:00 EST 1988}
}