Role of DNA polymerase eta in the UV mutation spectrum in human cells.
In humans, inactivation of the DNA polymerase eta gene (pol eta) results in sunlight sensitivity and causes the cancer-prone xeroderma pigmentosum variant syndrome (XP-V). Cells from XP-V individuals have a reduced capacity to replicate UV-damaged DNA and show hypermutability after UV exposure. Biochemical assays have demonstrated the ability of pol eta to bypass cis-syn-cyclobutane thymine dimers, the most common lesion generated in DNA by UV. In most cases, this bypass is error-free. To determine the actual requirement of pol eta in vivo, XP-V cells (XP30RO) were complemented by the wild type pol eta gene. We have used two pol eta-corrected clones to study the in vivo characteristics of mutations produced by DNA polymerases during DNA synthesis of UV-irradiated shuttle vectors transfected into human host cells, which had or had not been exposed previously to UV radiation. The functional complementation of XP-V cells by pol eta reduced the mutation frequencies both at CG and TA base pairs and restored UV mutagenesis to a normal level. UV irradiation of host cells prior to transfection strongly increased the mutation frequency in undamaged vectors and, in addition, especially in the pol eta-deficient XP30RO cells at 5'-TT sites in UV-irradiated plasmids. These results clearly show the protective role of pol eta against UV-induced lesions and the activation by UV of pol eta-independent mutagenic processes.
Stary
Anne
A
Laboratory of Genetic Instability and Cancer, UPR 2169 CNRS, Institut Gustave Roussy, 94805 Villejuif, France. stary@igr.fr
Kannouche
Patricia
P
Lehmann
Alan R
AR
Sarasin
Alain
A
D001483:Base Sequence; D002110:Caffeine / Q000494:pharmacology; D002453:Cell Cycle; D002460:Cell Line; D002461:Cell Line, Transformed; D002470:Cell Survival; D004249:DNA Damage; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D017353:Gene Deletion; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009154:Mutation; D010957:Plasmids; D011740:Pyrimidine Dimers; D017422:Sequence Analysis, DNA; D013539:Simian virus 40; D014162:Transfection; D014466:Ultraviolet Rays / Q000009:adverse effects*; D014983:Xeroderma Pigmentosum / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics; D000099270:Y-Family DNA Polymerases
D002110:Caffeine; D004259:DNA-Directed DNA Polymerase; D011740:Pyrimidine Dimers; D001616:beta-Galactosidase; C112338:Rad30 protein; D000099270:Y-Family DNA Polymerases