Frequency of intrachromosomal homologous recombination induced by UV radiation in normally repairing and excision repair-deficient human cells.
To investigate the role of DNA damage and nucleotide excision repair in intrachromosomal homologous recombination, a plasmid containing duplicated copies of the gene coding for hygromycin resistance was introduced into the genome of a repair-proficient human cell line, KMST-6, and two repair-deficient lines, XP2OS(SV) from xeroderma pigmentosum complementation group A and XP2YO(SV) from complementation group F. Neither hygromycin-resistance gene codes for a functional enzyme because each contains an insertion/deletion mutation at a unique site, but recombination between the two defective genes can yield hygromycin-resistant cells. The rates of spontaneous recombination in normal and xeroderma pigmentosum cell strains containing the recombination substrate were found to be similar. The frequency of UV-induced recombination was determined for three of these cell strains. At low doses, the group A cell strain and the group F cell strain showed a significant increase in frequency of recombinants. The repair-proficient cell strain required 10- to 20-fold higher doses of UV to exhibit comparable increases in frequency of recombinants. These results suggest that unexcised DNA damage, rather than the excision repair process per se, stimulates such recombination.
Tsujimura
T
T
Department of Microbiology, Michigan State University, East Lansing 48824-1316.
Maher
V M
VM
Godwin
A R
AR
Liskay
R M
RM
McCormick
J J
JJ
D015139:Blotting, Southern; D002460:Cell Line; D002470:Cell Survival / Q000528:radiation effects; D002877:Chromosomes, Human / Q000528:radiation effects*; D004260:DNA Repair; D004307:Dose-Response Relationship, Radiation; D005814:Genes, Viral; D006801:Humans; D009154:Mutation; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic / Q000528:radiation effects*; D015183:Restriction Mapping; D018139:Simplexvirus / Q000201:enzymology / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics; D014162:Transfection; D014466:Ultraviolet Rays; D015678:Viral Structural Proteins / Q000235:genetics; D014983:Xeroderma Pigmentosum / Q000235:genetics
D015678:Viral Structural Proteins; D013937:Thymidine Kinase