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PMID: 6393125 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A separate editing exonuclease for DNA replication: the epsilon subunit of Escherichia coli DNA polymerase III holoenzyme.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 81 ·No. 24 ·1984-12-00 ·Pages 7747-51

Scheuermann RH, Echols H

Abstract

DNA polymerase III (polIII) holoenzyme of Escherichia coli has 3'----5' exonuclease ("editing") activity in addition to its polymerase activity, a property shared by other prokaryotic DNA polymerases. The polymerization activity is carried by the large alpha subunit, the product of the dnaE gene. Mutations affecting the fidelity of DNA replication in vivo and the activity of 3'----5' exonuclease assayed in vitro are found in the dnaQ gene, which specifies the epsilon subunit. To determine whether epsilon carries the 3'----5' exonuclease activity, we have used an overproduction protocol to purify epsilon separately from the other subunits of polIII holoenzyme. We find that epsilon has 3'----5' exonuclease activity indistinguishable from that of polIII core, the subassembly of polIII holoenzyme consisting of the alpha, epsilon, and theta subunits. We conclude that the editing and polymerization activities of polIII holoenzyme reside on distinct subunits, in contrast to DNA polymerase I of E. coli and DNA polymerase of phage T4. This functional separation may provide for regulation of exonucleolytic editing independently of polymerization, allowing cellular control of replication fidelity.

MeSH Terms
DNA Polymerase III/isolation & purification,metabolism DNA Replication DNA-Directed DNA Polymerase/metabolism Escherichia coli/enzymology,genetics Kinetics Macromolecular Substances Molecular Weight Plasmids
Chemicals
Macromolecular Substances DNA Polymerase III DNA-Directed DNA Polymerase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Scheuermann R H
Echols H
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.81.24.7747
Published
1984-12-00
Pages
7747-51
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC392229
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.81.24.7747 PMC Full Text View journal details
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