Document Detail


Fidelity of DNA polymerase I and the DNA polymerase I-DNA primase complex from Saccharomyces cerevisiae.
MedLine Citation:
PMID:  2555694     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We have determined the fidelity of DNA synthesis by DNA polymerase I (yPol I) from Saccharomyces cerevisiae. To determine whether subunits other than the polymerase catalytic subunit influence fidelity, we measured the accuracy of yPol I purified by conventional procedures, which yields DNA polymerase with a partially proteolyzed catalytic subunit and no associated primase activity, and that of yPol I purified by immunoaffinity chromatography, which yields polymerase having a single high-molecular-weight species of the catalytic subunit, as well as three additional polypeptides and DNA primase activity. In assays that score polymerase errors within the lacZ alpha-complementation gene in M13mp2 DNA, yPol I and the yPol I-primase complex produced single-base substitutions, single-base frameshifts, and larger deletions. For specific errors and template positions, the two forms of polymerase exhibited differences in fidelity that could be as large as 10-fold. Nevertheless, results for the overall error frequency and the spectrum of errors suggest that the yPol I-DNA primase complex is not highly accurate and that, just as for the polymerase alone, its fidelity is not sufficient to account for a low spontaneous mutation rate in vivo. The specificity data also suggest models to explain -1 base frameshifts in nonrepeated sequences and certain complex deletions by a direct repeat mechanism involving aberrant loop-back synthesis.
Authors:
T A Kunkel; R K Hamatake; J Motto-Fox; M P Fitzgerald; A Sugino
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Molecular and cellular biology     Volume:  9     ISSN:  0270-7306     ISO Abbreviation:  Mol. Cell. Biol.     Publication Date:  1989 Oct 
Date Detail:
Created Date:  1989-12-29     Completed Date:  1989-12-29     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  8109087     Medline TA:  Mol Cell Biol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  4447-58     Citation Subset:  IM    
Affiliation:
Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709.
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MeSH Terms
Descriptor/Qualifier:
Base Sequence
Chromatography, Affinity
DNA Polymerase I / isolation & purification,  metabolism*
DNA Primase
DNA Repair / physiology*
DNA Replication / physiology*
Exonucleases / analysis
Immunosorbents
Models, Genetic
Molecular Sequence Data
Multienzyme Complexes / isolation & purification,  metabolism
Mutation
RNA Nucleotidyltransferases / metabolism*
Saccharomyces cerevisiae / enzymology*,  genetics,  physiology
Templates, Genetic
Chemical
Reg. No./Substance:
0/Immunosorbents; 0/Multienzyme Complexes; EC 2.7.7.-/DNA Polymerase I; EC 2.7.7.-/DNA Primase; EC 2.7.7.-/RNA Nucleotidyltransferases; EC 3.1.-/Exonucleases
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