Mutations associated with base excision repair deficiency and methylation-induced genotoxic stress

Proc Natl Acad Sci U S A. 2002 May 14;99(10):6860-5. doi: 10.1073/pnas.092662499. Epub 2002 Apr 30.

Abstract

The long-term effect of exposure to DNA alkylating agents is entwined with the cell's genetic capacity for DNA repair and appropriate DNA damage responses. A unique combination of environmental exposure and deficiency in these responses can lead to genomic instability; this "gene-environment interaction" paradigm is a theme for research on chronic disease etiology. In the present study, we used mouse embryonic fibroblasts with a gene deletion in the base excision repair (BER) enzymes DNA beta-polymerase (beta-pol) and alkyladenine DNA glycosylase (AAG), along with exposure to methyl methanesulfonate (MMS) to study mutagenesis as a function of a particular gene-environment interaction. The beta-pol null cells, defective in BER, exhibit a modest increase in spontaneous mutagenesis compared with wild-type cells. MMS exposure increases mutant frequency in beta-pol null cells, but not in isogenic wild-type cells; UV light exposure or N-methyl-N'-nitro-N-nitrosoguanidine exposure increases mutant frequency similarly in both cell lines. The MMS-induced increase in mutant frequency in beta-pol null cells appears to be caused by DNA lesions that are AAG substrates, because overexpression of AAG in beta-pol null cells eliminates the effect. In contrast, beta-pol/AAG double null cells are slightly more mutable than the beta-pol null cells after MMS exposure. These results illustrate that BER plays a role in protecting mouse embryonic fibroblast cells against methylation-induced mutations and characterize the effect of a particular combination of BER gene defect and environmental exposure.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alkylating Agents / pharmacology
  • Animals
  • Bacteriophage lambda / genetics
  • Base Pairing*
  • Binding Sites
  • Carbon-Oxygen Lyases / metabolism
  • Cells, Cultured
  • DNA Damage*
  • DNA Glycosylases*
  • DNA Methylation*
  • DNA Polymerase beta / genetics
  • DNA Polymerase beta / metabolism*
  • DNA Repair*
  • DNA-(Apurinic or Apyrimidinic Site) Lyase
  • Genes, Viral
  • Mesylates / pharmacology
  • Methylnitronitrosoguanidine / pharmacology
  • Mice
  • Mice, Knockout
  • Mutagenesis
  • Mutagens / pharmacology
  • N-Glycosyl Hydrolases / genetics
  • N-Glycosyl Hydrolases / metabolism*
  • Transcription Factors / genetics
  • Viral Proteins

Substances

  • Alkylating Agents
  • Mesylates
  • Mutagens
  • Transcription Factors
  • Viral Proteins
  • cII protein, bacteriophage lambda
  • methanesulfonic acid
  • Methylnitronitrosoguanidine
  • DNA Polymerase beta
  • 3-methyladenine-DNA glycosylase
  • DNA Glycosylases
  • N-Glycosyl Hydrolases
  • Carbon-Oxygen Lyases
  • DNA-(Apurinic or Apyrimidinic Site) Lyase