TY - JOUR
T1 - Phenotypic heterogeneity in nucleotide excision repair mutants of rodent complementation groups 1 and 4
AU - Busch, David B.
AU - Van Vuuren, Hanneke
AU - De Wit, Jan
AU - Collins, Andrew
AU - Zdzienicka, Małgorzata Z.
AU - Mitchell, David L.
AU - Brookman, Kerry W.
AU - Stefanini, Miria
AU - Riboni, Roberta
AU - Thompson, Larry H.
AU - Albert, Roberta Bliss
AU - Van Gool, Alain J.
AU - Hoeijmakers, Jan
N1 - Funding Information:
The work reported here was supported by the Armed Forces Institute of Pathology, APC UBDH; the American Registry of Pathology; Dutch Cancer Society Grant IKW 90-03, Cancer Research Campaign; Scottish Office Agriculture and Fisheries Department; the National Institutes of Health (Grants GM22021, CA04484, and RR00961); the Dutch Scientific Organization, area Medical Sciences (Project 901-501-113, 093) and the Section of Nucleic Acids and HSFP; and the National Research Council of Italy (C.N.R. Target Project `Ingegneria Genetica') and performed under the auspices of the US Department of Energy by Lawrence Berkeley Laboratory Grant 7134700, and Lawrence Livermore National Laboratory Contract W-7405-ENG-48. We appreciate the advice and assistance of Donald A. Glaser and technical support offered by Jerry Adams, Hazel Bell, John L. Couch, Ruth Ford, Carol Greiner, Hanneke Kool, Kathy Lewis, Alex Para, Helen Stouffer, and Lisa Wills. The opinions or assertions stated herein are the private views of the authors, and are not to be construed as representing the views of the American Registry of Pathology, the Armed Forces Institute of Pathology, the Department of the Army, the Department of Environmental and Toxicologic Pathology, Lawrence Livermore National Laboratory, the US Department of Defense, or the US Department of Energy.
PY - 1997/3/12
Y1 - 1997/3/12
N2 - 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.
AB - 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.
KW - Allele
KW - Chinese hamster ovary (CHO)
KW - DNA repair
KW - Mitomycin C
KW - Mutant
KW - Ultraviolet light
UR - http://www.scopus.com/inward/record.url?scp=17744411864&partnerID=8YFLogxK
U2 - 10.1016/S0921-8777(96)00048-1
DO - 10.1016/S0921-8777(96)00048-1
M3 - Article
C2 - 9088342
AN - SCOPUS:17744411864
SN - 0921-8777
VL - 383
SP - 91
EP - 106
JO - Mutation Research-DNA Repair
JF - Mutation Research-DNA Repair
IS - 2
ER -