Mutagenesis and repair induced by the DNA advanced glycation end product N2-1-(carboxyethyl)-2'-deoxyguanosine in human cells

Biochemistry. 2011 Mar 29;50(12):2321-9. doi: 10.1021/bi101933p. Epub 2011 Feb 28.


Glycation of biopolymers by glucose-derived α-oxo-aldehydes such as methylglyoxal (MG) is believed to play a major role in the complex pathologies associated with diabetes and metabolic disease. In contrast to the extensive literature detailing the formation and physiological consequences of protein glycation, there is little information about the corresponding phenomenon for DNA. To assess the potential contribution of DNA glycation to genetic instability, we prepared shuttle vectors containing defined levels of the DNA glycation adduct N(2)-(1-carboxyethyl)-2'-deoxyguanosine (CEdG) and transfected them into isogenic human fibroblasts that differed solely in the capacity to conduct nucleotide excision repair (NER). In the NER-compromised fibroblasts, the induced mutation frequencies increased up to 18-fold relative to background over a range of ∼10-1400 CEdG adducts/10(5) dG, whereas the same substrates transfected into NER-competent cells induced a response that was 5-fold over background at the highest adduct density. The positive linear correlation (R(2) = 0.998) of mutation frequency with increasing CEdG level in NER-defective cells suggested that NER was the primary if not exclusive mechanism for repair of this adduct in human fibroblasts. Consistent with predictions from biochemical studies using CEdG-substituted oligonucleotides, guanine transversions were the predominant mutation resulting from replication of MG-modified plasmids. At high CEdG levels, significant increases in the number of AT → GC transitions were observed exclusively in NER-competent cells (P < 0.0001). This suggested the involvement of an NER-dependent mutagenic process in response to critical levels of DNA damage, possibly mediated by error-prone Y-family polymerases.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA Adducts / genetics*
  • DNA Adducts / metabolism*
  • DNA Repair*
  • Deoxyguanosine / analogs & derivatives*
  • Deoxyguanosine / metabolism
  • Fibroblasts / metabolism*
  • Genes, Suppressor
  • Glycation End Products, Advanced / metabolism*
  • Humans
  • Mutagenesis*
  • Mutation
  • Plasmids / genetics
  • RNA, Transfer / genetics


  • DNA Adducts
  • Glycation End Products, Advanced
  • N(2)-(1-carboxyethyl)deoxyguanosine
  • supF tRNA
  • RNA, Transfer
  • Deoxyguanosine