Direct measurement of the poliovirus RNA polymerase error frequency in vitro

Journal Article · Mon Feb 01 00:00:00 EST 1988 · Journal of Virology; (USA)

OSTI ID:6525520

Ward, C D; Stokes, M A.M.; Flanegan, J B ^[1]

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1. Univ. of Florida College of Medicine, Gainesville (USA)

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.

• OSTI ID: 6525520
• Journal Information: Journal of Virology; (USA), Vol. 62:2; ISSN 0022-538X
• Country of Publication: United States
• Language: English

Related Subjects

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