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. 1993 May 15;90(10):4626–4630. doi: 10.1073/pnas.90.10.4626

Detection and characterization of mammalian DNA polymerase beta mutants by functional complementation in Escherichia coli.

J B Sweasy 1, L A Loeb 1
PMCID: PMC46565  PMID: 8506308

Abstract

We have designed and utilized a bacterial complementation system to identify and characterize mammalian DNA polymerase beta mutants. In this complementation system, wild-type rat DNA polymerase beta replaces both the replicative and repair functions of DNA polymerase I in the Escherichia coli recA718 polA12 double mutant; our 263 DNA polymerase beta mutants replace E. coli polymerase I less efficiently or not at all. Of the 10 mutants that have been shown to contain DNA sequence alterations, 2 exhibit a split phenotype with respect to complementation of the growth defect and methylmethanesulfonate sensitivity of the double mutant; one is a null mutant. The mutants possessing a split phenotype contain amino acid residue alterations within a putative nucleotide binding site of DNA polymerase beta. This approach for the isolation and evaluation of mutants of a mammalian DNA polymerase in E. coli may ultimately lead to a better understanding of the mechanism of action of this enzyme and to precisely defining its role in vertebrate cells.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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