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Archaeal DNA polymerase D but not DNA polymerase B is required for genome replication in Thermococcus kodakarensis

Lookup NU author(s): Dr Tom Richardson, Professor Bernard Connolly

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Abstract

Three evolutionarily distinct families of replicative DNA polymerases, designated Pol B, Pol C and Pol D, have been identified. Members of the Pol B family are present in all three Domains of life whereas Pol C exists only in Bacteria and Pol D only in Archaea. Pol B enzymes replicate eukaryotic chromosomal DNA, and as members of the Pol B family are present in all Archaea, it has been assumed that Pol B enzymes also replicate archaeal genomes. Here we report the construction of Thermococcus kodakarensis strains with mutations that delete or inactivate key functions of Pol B. T. kodakarensis strains lacking Pol B had no detectable loss in viability, no growth defects nor changes in spontaneous mutation frequency, but have increased sensitivity to UV irradiation. In contrast, we were unable to introduce mutations that inactivated either of the genes encoding the two subunits of Pol D. The results reported establish that Pol D is sufficient for viability and genome replication in T. kodakarensis and argue that Pol D rather than Pol B is likely the replicative DNA polymerase in this Archaeon. The majority of Archaea contain Pol D and, as discussed, if Pol D is the predominant replicative polymerase in Archaea, this profoundly impacts hypotheses for the origin(s), evolution and distribution of the different DNA replication enzymes and systems now employed in the three Domains of life.


Publication metadata

Author(s): Cubonova L, Richardson T, Burkhart BW, Kelma Z, Connolly BA, Reeve JN, Santangelo TJ

Publication type: Article

Publication status: Published

Journal: Journal of Bacteriology

Year: 2013

Volume: 195

Issue: 10

Pages: 2322-2328

Print publication date: 01/05/2013

Online publication date: 15/03/2013

Acceptance date: 04/03/2013

ISSN (print): 0021-9193

ISSN (electronic): 1098-5530

Publisher: American Society for Microbiology

URL: http://dx.doi.org/10.1128/JB.02037-12

DOI: 10.1128/JB.02037-12

PubMed id: 23504010


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