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

Joann B. Sweasy, Lawrence A. Loeb

Research output: Contribution to journalArticlepeer-review

44 Scopus citations

Abstract

We have designed and utilized a bacterial complementation system to identify and characterize mammalian DNA polymerase β mutants. In this complementation system, wild-type rat DNA polymerase β replaces both the replicative and repair functions of DNA polymerase I in the Escherichia coli recA718 polA12 double mutant; our 263 DNA polymerase β 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 β. 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.

Original languageEnglish (US)
Pages (from-to)4626-4630
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number10
DOIs
StatePublished - May 15 1993
Externally publishedYes

Keywords

  • DNA replication
  • Mutagenesis

ASJC Scopus subject areas

  • General

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