Involvement of DnaE, the second replicative DNA polymerase from Bacillus subtilis, in DNA mutagenesis

Le Chatelier, Emmanuelle, Becherel, Olivier J., d'Alencon, Emmanuelle, Canceill, Danielle, Ehrlich, S. Dusko, Fuchs, Robert P. P. and Janniere, Laurent (2004) Involvement of DnaE, the second replicative DNA polymerase from Bacillus subtilis, in DNA mutagenesis. Journal of Biological Chemistry, 279 3: 1757-1767. doi:10.1074/jbc.M310719200

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Author Le Chatelier, Emmanuelle
Becherel, Olivier J.
d'Alencon, Emmanuelle
Canceill, Danielle
Ehrlich, S. Dusko
Fuchs, Robert P. P.
Janniere, Laurent
Title Involvement of DnaE, the second replicative DNA polymerase from Bacillus subtilis, in DNA mutagenesis
Formatted title
Involvement of DnaE, the second replicative DNA polymerase from Bacillus subtilis, in DNA mutagenesis
Journal name Journal of Biological Chemistry   Check publisher's open access policy
ISSN 0021-9258
1083-351X
Publication date 2004-01-16
Sub-type Article (original research)
DOI 10.1074/jbc.M310719200
Open Access Status File (Publisher version)
Volume 279
Issue 3
Start page 1757
End page 1767
Total pages 11
Place of publication Bethesda, MD, United States
Publisher American Society for Biochemistry and Molecular Biology
Language eng
Formatted abstract
In a large group of organisms including low G + C bacteria and eukaryotic cells, DNA synthesis at the replication fork strictly requires two distinct replicative DNA polymerases. These are designated pol C and DnaE in Bacillus subtilis. We recently proposed that DnaE might be preferentially involved in lagging strand synthesis, whereas pol C would mainly carry out leading strand synthesis. The biochemical analysis of DnaE reported here is consistent with its postulated function, as it is a highly potent enzyme, replicating as fast as 240 nucleotides/s, and stalling for more than 30 s when encountering annealed 5′-DNA end. DnaE is devoid of 3′ → 5′-proofreading exonuclease activity and has a low processivity (1-75 nucleotides), suggesting that it requires additional factors to fulfill its role in replication. Interestingly, we found that (i) DnaE is SOS-inducible; (ii) variation in DnaE or pol C concentration has no effect on spontaneous mutagenesis; (iii) depletion of pol C or DnaE prevents UV-induced mutagenesis; and (iv) purified DnaE has a rather relaxed active site as it can bypass lesions that generally block other replicative polymerases. These results suggest that DnaE and possibly pol C have a function in DNA repair/mutagenesis, in addition to their role in DNA replication.
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: School of Chemistry and Molecular Biosciences
 
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