UV mutagenesis, cytotoxicity and split-dose recovery in a human-CHO cell hybrid having intermediate (6-4) photoproduct repair.
Somatic cell hybrids constructed between UV-hypersensitive Chinese hamster ovary cell line UV20 and human lymphocytes were used to examine the influence of a human DNA repair gene, ERCC1, on UV photoproduct repair, mutability at several drug-resistance loci, UV cytotoxicity and UV split-dose recovery. In hybrid cell line 20HL21-4, which contains human chromosome 19, UV-induced mutagenesis at the APRT, HPRT and Na+/K+-ATPase loci was comparable to that in repair-proficient CHO AA8 cells, whereas cell line 20HL21-7, a reduced human-CHO hybrid not containing human chromosome 19, exhibited a hypermutable phenotype at all 3 loci indistinguishable from that of UV20 cells. The response of 20HL21-4 cells to UV cytotoxicity reflected substantial but incomplete restoration of wild-type UV cytotoxic response, whereas responses of UV20 and 20HL21-7 cell lines to UV cytotoxicity were essentially the same, reflecting several-fold UV hypersensitivity. Repair of UV-induced (5-6) cyclobutane dimers and (6-4) photoproducts was examined by radioimmunoassay; (6-4) photoproduct repair was deficient in UV20 and 20HL21-7 cell lines, and intermediate in 20HL21-4 cells relative to wild-type CHO AA8 cells. UV split-dose recovery in 20HL21-4 cells was also intermediate relative to AA8 cells. These results show that the human ERCC1 gene on chromosome 19 is responsible for substantial restoration of UV survival and mutation responses in repair-deficient UV20 cells, but only partially restores (6-4) UV photoproduct repair and UV split-dose recovery.
Nairn
R S
RS
University of Texas, M.D. Anderson Cancer Center, Department of Experimental Carcinogenesis, Smithville 78957.
Mitchell
D L
DL
Adair
G M
GM
Thompson
L H
LH
Siciliano
M J
MJ
Humphrey
R M
RM
D000228:Adenine Phosphoribosyltransferase / Q000235:genetics; D000818:Animals; D002470:Cell Survival / Q000528:radiation effects*; D002888:Chromosomes, Human, Pair 19; D006224:Cricetinae; D004260:DNA Repair; D004307:Dose-Response Relationship, Radiation; D006801:Humans; D006822:Hybrid Cells / Q000528:radiation effects; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D009154:Mutation / Q000528:radiation effects*; D011740:Pyrimidine Dimers; D000254:Sodium-Potassium-Exchanging ATPase / Q000235:genetics
D011740:Pyrimidine Dimers; D000228:Adenine Phosphoribosyltransferase; D007041:Hypoxanthine Phosphoribosyltransferase; D000254:Sodium-Potassium-Exchanging ATPase