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PMID: 3001347 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Direct method for quantitation of extreme polymerase error frequencies at selected single base sites in viral RNA.

Journal of virology ·Vol. 57 ·No. 1 ·1986-01-00 ·Pages 219-28

Steinhauer DA, Holland JJ

Abstract

Methods are described which allow direct quantitation and sequence analysis of base substitution levels at predetermined single nucleotide positions in cloned pools of an RNA virus genome or in its RNA transcripts in vitro. Base substitution frequencies for vesicular stomatitis virus (VSV) at one highly conserved site examined were reproducible and extremely high, averaging between 10(-4) and 4 X 10(-4) substitutions per base incorporated at this single site. If polymerase error frequencies averaged as high at all other sites in the 11-kilobase VSV genome, then every member of a cloned VSV population would differ from most other genomes in that clone at a number of different nucleotide positions. The preservation of a consensus sequence in such variable RNA virus genomes then could only result from strong biological selection (in a single host or multihost environment) for the most fit and competitive representatives of extremely heterogeneous virus populations.

MeSH Terms
Base Sequence DNA-Directed RNA Polymerases/physiology Defective Viruses/genetics Electrophoresis, Polyacrylamide Gel Methods Mutation Oligonucleotides/analysis RNA Nucleotidyltransferases/physiology RNA, Messenger/analysis RNA, Viral/analysis RNA-Dependent RNA Polymerase/physiology Selection, Genetic Vesicular stomatitis Indiana virus/genetics
Chemicals
Oligonucleotides RNA, Messenger RNA, Viral RNA Nucleotidyltransferases RNA-Dependent RNA Polymerase DNA-Directed RNA Polymerases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Steinhauer D A
Holland J J
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.57.1.219-228.1986
Published
1986-01-00
Pages
219-28
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC252718
Subset
IM
Grants
NIAID NIH HHS · AI14627 · United States
External Links
PubMed source DOI: 10.1128/JVI.57.1.219-228.1986 PMC Full Text View journal details
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