Molecular nature of ultraviolet B light-induced deletions in the murine epidermis

Cancer Res. 2001 May 15;61(10):3913-8.

Abstract

Depletion of the stratospheric ozone layer leads to an increase in ambient UV loads, which are expected to raise skin cancer incidences. Tumor development in the skin could be a multistep process in which various genetic alterations, such as point mutations and deletions, occur successively. Here, we demonstrate that UVB irradiation efficiently induces deletions in the epidermis using a novel transgenic mouse, gpt delta. In this mouse model, deletions in lambda DNA integrated in the chromosome are preferentially selected as Spi(-) (sensitive to P2 interference) phages, which can then be subjected to molecular analysis. The mice were exposed to UVB at single doses of 0.3, 0.5, 1.0, 1.5, and 2.0 kJ/m(2). After 4 weeks, lambda phage was rescued from the genomic DNA of the epidermis by in vitro packaging reactions. The mutant frequencies of Spi(-) with large deletions in the epidermis increased >15-fold at a UVB dose of 0.5 kJ/m(2) over the control. Molecular sizes of most of the large deletions were >1000 bp. More than one-half of the large deletions occurred between short direct-repeat sequences from 1 to 6 bp, and the remainder had flush ends. In the unirradiated mouse, almost all of the Spi(-) mutants were 1-bp frameshifts in runs of identical bases. These results suggest that UVB irradiation induces deletions in the murine epidermis, and most of the deletions are generated through end-joining of double strand breaks in DNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Bacteriophage lambda / genetics
  • Base Sequence
  • DNA, Viral / genetics
  • DNA, Viral / radiation effects
  • Dose-Response Relationship, Radiation
  • Epidermis / physiology
  • Epidermis / radiation effects*
  • Escherichia coli / genetics
  • Escherichia coli / virology
  • Escherichia coli Proteins
  • Female
  • Frameshift Mutation
  • Male
  • Mice
  • Mice, Transgenic
  • Pentosyltransferases
  • Proteins*
  • Sequence Deletion*
  • Skin Neoplasms / etiology
  • Skin Neoplasms / genetics
  • Ultraviolet Rays / adverse effects*

Substances

  • Bacterial Proteins
  • DNA, Viral
  • Escherichia coli Proteins
  • Proteins
  • Pentosyltransferases
  • Gpt protein, E coli