Defining the function of xeroderma pigmentosum group F protein in psoralen interstrand cross-link-mediated DNA repair and mutagenesis

Biochem J. 2004 Apr 1;379(Pt 1):71-8. doi: 10.1042/BJ20031143.

Abstract

Many commonly used drugs, such as psoralen and cisplatin, can generate a very unique type of DNA damage, namely ICL (interstrand cross-link). An ICL can severely block DNA replication and transcription and cause programmed cell death. The molecular mechanism of repairing the ICL damage has not been well established. We have studied the role of XPF (xeroderma pigmentosum group F) protein in psoralen-induced ICL-mediated DNA repair and mutagenesis. The results obtained from our mutagenesis studies revealed a very similar mutation frequency in both human normal fibroblast cells and XPF cells. The mutation spectra generated in both cells, however, were very different: most of the mutations generated in the normal fibroblast cells were T167-->A transversions, whereas most of the mutations generated in the XPF cells were T167-->G transversions. When a wild-type XPF gene cDNA was stably transfected into the XPF cells, the T167-->A mutations were increased and the T167-->G mutations were decreased. We also determined the DNA repair capability of the XPF cells using both the host-cell reactivation and the in vitro DNA repair assays. The results obtained from the host-cell reactivation experiments revealed an effective reactivation of a luciferase reporter gene from the psoralen-damaged plasmid in the XPF cells. The results obtained from the in vitro DNA repair experiments demonstrated that the XPF nuclear extract is normal in introducing dual incisions during the nucleotide excision repair process. These results suggest that the XPF protein has important roles in the psoralen ICL-mediated DNA repair and mutagenesis.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Sequence
  • Cells, Cultured / drug effects
  • Cells, Cultured / metabolism
  • Cells, Cultured / radiation effects
  • Cross-Linking Reagents / pharmacology*
  • DNA Adducts / metabolism
  • DNA Damage*
  • DNA Repair / genetics
  • DNA Repair / physiology*
  • DNA, Complementary / genetics
  • DNA, Recombinant / drug effects*
  • DNA, Recombinant / genetics
  • DNA, Recombinant / radiation effects
  • DNA-Binding Proteins / deficiency
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / physiology*
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • Fibroblasts / radiation effects
  • Genes, Reporter
  • Genes, Suppressor
  • Genes, Synthetic / drug effects
  • Genes, Synthetic / radiation effects
  • Humans
  • Luciferases / biosynthesis
  • Luciferases / genetics
  • Molecular Sequence Data
  • Mutagenesis*
  • Mutagenesis, Site-Directed
  • Oligonucleotides / pharmacology*
  • Photochemistry
  • Plasmids / drug effects
  • Plasmids / radiation effects
  • RNA, Transfer / genetics
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics
  • Transfection
  • Trioxsalen / analogs & derivatives
  • Trioxsalen / pharmacology*
  • Ultraviolet Rays
  • Xeroderma Pigmentosum / genetics*
  • Xeroderma Pigmentosum / metabolism
  • Xeroderma Pigmentosum / pathology

Substances

  • Cross-Linking Reagents
  • DNA Adducts
  • DNA, Complementary
  • DNA, Recombinant
  • DNA-Binding Proteins
  • Oligonucleotides
  • Recombinant Fusion Proteins
  • psoAG10 compound
  • supF tRNA
  • xeroderma pigmentosum group F protein
  • RNA, Transfer
  • Luciferases
  • Trioxsalen