| Fidelity of DNA polymerase I and the DNA polymerase I-DNA primase complex from Saccharomyces cerevisiae. | |
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MedLine Citation:
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PMID: 2555694 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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T A Kunkel; R K Hamatake; J Motto-Fox; M P Fitzgerald; A Sugino |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Molecular and cellular biology Volume: 9 ISSN: 0270-7306 ISO Abbreviation: Mol. Cell. Biol. Publication Date: 1989 Oct |
Date Detail:
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Created Date: 1989-12-29 Completed Date: 1989-12-29 Revised Date: 2009-11-18 |
Medline Journal Info:
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Nlm Unique ID: 8109087 Medline TA: Mol Cell Biol Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 4447-58 Citation Subset: IM |
Affiliation:
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Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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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:
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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 |
| Comments/Corrections | |
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