UV radiation induces delayed hyperrecombination associated with hypermutation in human cells.
Ionizing radiation induces delayed genomic instability in human cells, including chromosomal abnormalities and hyperrecombination. Here, we investigate delayed genome instability of cells exposed to UV radiation. We examined homologous recombination-mediated reactivation of a green fluorescent protein (GFP) gene in p53-proficient human cells. We observed an approximately 5-fold enhancement of delayed hyperrecombination (DHR) among cells surviving a low dose of UV-C (5 J/m2), revealed as mixed GFP+/- colonies. UV-B did not induce DHR at an equitoxic (75 J/m2) dose or a higher dose (150 J/m2). UV is known to induce delayed hypermutation associated with increased oxidative stress. We found that hypoxanthine phosphoribosyltransferase (HPRT) mutation frequencies were approximately 5-fold higher in strains derived from GFP+/- (DHR) colonies than in strains in which recombination was directly induced by UV (GFP+ colonies). To determine whether hypermutation was directly caused by hyperrecombination, we analyzed hprt mutation spectra. Large-scale alterations reflecting large deletions and insertions were observed in 25% of GFP+ strains, and most mutants had a single change in HPRT. In striking contrast, all mutations arising in the hypermutable GFP+/- strains were small (1- to 2-base) changes, including substitutions, deletions, and insertions (reminiscent of mutagenesis from oxidative damage), and the majority were compound, with an average of four hprt mutations per mutant. The absence of large hprt deletions in DHR strains indicates that DHR does not cause hypermutation. We propose that UV-induced DHR and hypermutation result from a common source, namely, increased oxidative stress. These two forms of delayed genome instability may collaborate in skin cancer initiation and progression.
Durant
Stephen T
ST
Department of Molecular Genetics and Microbiology, University of New Mexico School of Medicine, Albuquerque, NM 87131, USA.
Paffett
Kimberly S
KS
Shrivastav
Meena
M
Timmins
Graham S
GS
Morgan
William F
WF
Nickoloff
Jac A
JA
D016923:Cell Death / Q000528:radiation effects; D002470:Cell Survival / Q000528:radiation effects; D005091:Exons / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D008954:Models, Biological; D016296:Mutagenesis / Q000235:genetics / Q000528:radiation effects*; D017354:Point Mutation / Q000235:genetics*; D011995:Recombination, Genetic / Q000235:genetics / Q000528:radiation effects*; D014407:Tumor Cells, Cultured; D014466:Ultraviolet Rays
D049452:Green Fluorescent Proteins; D007041:Hypoxanthine Phosphoribosyltransferase