Molecular nature of ultraviolet B light-induced deletions in the murine epidermis.
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.
Horiguchi
M
M
Division of Genetics and Mutagenesis, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan.
Masumura
K I
KI
Ikehata
H
H
Ono
T
T
Kanke
Y
Y
Nohmi
T
T
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D004279:DNA, Viral / Q000235:genetics / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D004817:Epidermis / Q000502:physiology / Q000528:radiation effects*; D004926:Escherichia coli / Q000235:genetics / Q000821:virology; D029968:Escherichia coli Proteins; D005260:Female; D016368:Frameshift Mutation; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D010430:Pentosyltransferases; D011506:Proteins; D017384:Sequence Deletion; D012878:Skin Neoplasms / Q000209:etiology / Q000235:genetics; D014466:Ultraviolet Rays / Q000009:adverse effects*
D001426:Bacterial Proteins; D004279:DNA, Viral; D029968:Escherichia coli Proteins; D011506:Proteins; D010430:Pentosyltransferases; C071962:Gpt protein, E coli