Mutation frequencies and spectra in DNA polymerase eta-deficient mice.
The low-fidelity polymerase eta (poleta) is required for bypass of UV-induced pyrimidine dimers inserting adenine nucleotides opposite these lesions. Mutations in the poleta gene are responsible for the genetic defect in xeroderma pigmentosum variant patients. To study if the lack of poleta significantly elevates spontaneous mutation frequency in various organs and tissues of the mouse, we crossed poleta-deficient mice with transgenic mice harboring a chromosomally integrated lacZ-plasmid reporter construct. In cultured embryonic fibroblasts from the lacZ-poleta(-/-) mice, 2.5 J/m(2) UV irradiation induced approximately 5-fold more mutations than in cells from lacZ control mice, in which an approximately 3-fold increase in mutation frequency was found compared with the normal level. Whereas untreated cells harbored mainly 1-bp deletions, UV induced both transitions and transversions, with the latter type more highly represented in the poleta-null cells than in the controls. No difference in mutation induction between the poleta-null cells and the wild-type cells was observed after treatment with N-ethyl-N-nitrosourea. Having shown the validity of the lacZ model to accurately identify poleta-associated mutagenesis, we then determined the mutant frequency at the lacZ locus in liver, spleen, and small intestine of 12-month-old animals. No differences were found between poleta-null, heterozygous, or littermate control mice. We conclude that the poleta defect is specific for UV damage and has no effect on in vivo mutagenesis in mice.
Busuttil
Rita A
RA
Buck Institute for Age Research, Novato, CA 94945, USA.
Lin
Qingcong
Q
Stambrook
Peter J
PJ
Kucherlapati
Raju
R
Vijg
Jan
J
D000818:Animals; D004259:DNA-Directed DNA Polymerase / Q000172:deficiency*; D005038:Ethylnitrosourea; D005347:Fibroblasts / Q000187:drug effects / Q000528:radiation effects; D007763:Lac Operon; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D010957:Plasmids / Q000235:genetics; D017354:Point Mutation; D014466:Ultraviolet Rays; D000099270:Y-Family DNA Polymerases
D004259:DNA-Directed DNA Polymerase; D005038:Ethylnitrosourea; C112338:Rad30 protein; D000099270:Y-Family DNA Polymerases