Phenotypic heterogeneity in nucleotide excision repair mutants of rodent complementation groups 1 and 4
Mutation Research - DNA Repair , Volume 383 - Issue 2 p. 91- 106
Rodent ultraviolet light (UV)-sensitive mutant cells in complementation groups (CGs) 1 and 4 normally are known for their extraordinary (~80-100 x) sensitivity to mitomycin C (MMC), although some CG1 mutants with reduced MMC sensitivity were previously reported (Stefanini et al. (1987) Cytotechnology 1, 91). We report here new CG1 and CG4 mutants with only 1.6-10 x wild-type MMC sensitivity despite low unscheduled DNA synthesis (UDS) levels. Mutant UV140, in UV CG4, has ~3.8 x the UV sensitivity of parental line AA8, ~1.6 x wild-type MMC sensitivity, wild-type X-ray and ethyl methanesulfonate (EMS) sensitivity, and is only slightly (~1.4 x) hypermutable to 8-azaadenine resistance by UV light, It has moderately decreased Incision of UV-damaged DNA, has moderately decreased removal of (6-4) photoproducts, and is profoundly deficient in UDS after UV. After UV, it shows abnormally decreased DNA synthesis and persistently decreased RNA synthesis. In addition a cell-free extract of this mutant displays strongly reduced nucleotide excision repair synthesis using DNA treated with N-acetoxy-acetyl-amino-fluorene (AAF). The extract selectively fails to complement extracts of group 1 and 4 mutants consistent with the notion that the affected proteins, ERCC1 and ERCC4, are part of the same complex and that mutations in one subunit also affect the other component. Mutant UV212 is a CG1 mutant with ~3.3 x wild-type UV and ~5-10 x wild-type MMC sensitivity, with profoundly deficient UDS and hypermutability (~5.8 x) by UV, Mutant UV201, probably in CG1, is only slightly (~1.5 x) UV-sensitive and has near wild-type (1.02 x) UV mutability. These unusual group 1 and 4 mutants demonstrate that the unique UV and MMC sensitivity phenotypes displayed by these groups can be separated and support the idea that they are the result of distinct repair functions of the corresponding ERCC1 and ERCC4 genes: nucleotide excision repair for UV lesions and a separate repair pathway for removal of interstrand crosslinks.
|, , , , ,|
|Mutation Research - DNA Repair|
|Organisation||Department of Molecular Genetics|
Busch, D.B, van Vuuren, H, de Wit, J, Collins, A, Zdzienicka, M.Z, Mitchell, D.L, … Hoeijmakers, J.H.J. (1997). Phenotypic heterogeneity in nucleotide excision repair mutants of rodent complementation groups 1 and 4. Mutation Research - DNA Repair, 383(2), 91–106. doi:10.1016/S0921-8777(96)00048-1