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PMID: 15470496 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Defining the position of the switches between replicative and bypass DNA polymerases.

The EMBO journal ·Vol. 23 ·No. 21 ·2004-10-27 ·Pages 4342-52

Fujii S, Fuchs RP

Abstract

Cells contain specialized DNA polymerases that are able to copy past lesions with an associated risk of generating mutations, the major cause of cancer. Here, we reconstitute translesion synthesis (TLS) using the replicative (Pol III) and major bypass (Pol V) DNA polymerases from Escherichia coli in the presence of accessory factors. When the replicative polymerase disconnects from the template in the vicinity of a lesion, Pol V binds the blocked replication intermediate and forms a stable complex by means of a dual interaction with the tip of the RecA filament and the beta-clamp, the processivity factor donated by the blocked Pol III holoenzyme. Both interactions are required to confer to Pol V the processivity that will allow it synthesize, in a single binding event, a TLS patch long enough to support further extension by Pol III. In the absence of these accessory factors, the patch synthesized by Pol V is too short, being degraded by the Pol III-associated exonuclease activity that senses the distortion induced by the lesion, thus leading to an aborted bypass process.

MeSH Terms
DNA Damage DNA Polymerase III/metabolism DNA Replication DNA, Single-Stranded/metabolism DNA-Directed DNA Polymerase/metabolism Escherichia coli/genetics,metabolism Escherichia coli Proteins/genetics,metabolism Macromolecular Substances Nucleic Acid Conformation Rec A Recombinases/metabolism
Chemicals
DNA, Single-Stranded Escherichia coli Proteins Macromolecular Substances Rec A Recombinases DNA Polymerase III DNA polymerase V, E coli DNA-Directed DNA Polymerase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Fujii Shingo
UPR 9003 du CNRS, Cancerogenese et Mutagenese Moleculaire et Structurale, ESBS Pole API, Blvd S Brant Strasbourg, 67400 Illkirch-Graffenstaden, France.
Fuchs Robert P
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1038/sj.emboj.7600438
Published
2004-10-27
Epub
2004-00-07
Pages
4342-52
Language
English
Region
England
NLM ID
8208664
PMCID
PMC524402
Subset
IM
External Links
PubMed source DOI: 10.1038/sj.emboj.7600438 PMC Full Text View journal details
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