Molecular cloning of a mammalian gene involved in the fixation of UV-induced mutations.
A mammalian DNA damage tolerance gene has been isolated by DNA transfection and cosmid rescue. Following cotransfection of mouse genomic DNA and pSV2neo into SVM, the UV hypersensitive mutant Indian muntjac cell line, clones with a 1.7 to 2.0-fold greater D37 value for UV killing were isolated. This trait was carried through three rounds of transfection. A neo gene and flanking sequences from a tertiary transfectant were cloned by cosmid rescue. The cosmid clone confers UV resistance to SVM and improves the ability of the cell to replicate UV damaged DNA. This replication appears to be error-prone; UV-induced 6-thioguanine-resistant mutants occur four to fivefold more frequently than in SVM or a wild-type Indian muntjac line. Thus, the gene isolated is not homologous to that defective in SVM. We believe that this is the first mammalian gene to be isolated that is directly involved in mutation fixation.
Bouffler
S D
SD
Department of Zoology, University of Cambridge, U.K.
Godfrey
D
D
Raman
M J
MJ
Musk
S R
SR
Johnson
R T
RT
D000818:Animals; D015139:Blotting, Southern; D002460:Cell Line; D003001:Cloning, Molecular; D004260:DNA Repair; D004261:DNA Replication; D003670:Deer; D005816:Genetic Complementation Test; D051379:Mice; D016296:Mutagenesis; D015183:Restriction Mapping; D012854:Sister Chromatid Exchange; D014162:Transfection