Phenotypic heterogeneity in nucleotide excision repair mutants of rodent complementation groups 1 and 4

David B. Busch, Hanneke Van Vuuren, Jan De Wit, Andrew Collins, Małgorzata Z. Zdzienicka, David L. Mitchell, Kerry W. Brookman, Miria Stefanini, Roberta Riboni, Larry H. Thompson, Roberta Bliss Albert, Alain J. Van Gool, Jan Hoeijmakers

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)91-106
Number of pages16
JournalMutation Research - DNA Repair
Volume383
Issue number2
DOIs
Publication statusPublished - Mar 12 1997

Fingerprint

Ultraviolet Rays
DNA Repair
Rodentia
Repair
Nucleotides
Mitomycin
DNA
Ethyl Methanesulfonate
Photophobia
Photosensitivity
Cell Extracts
Genes
Display devices
X-Rays
RNA

Keywords

  • Allele
  • Chinese hamster ovary (CHO)
  • DNA repair
  • Mitomycin C
  • Mutant
  • Ultraviolet light

ASJC Scopus subject areas

  • Toxicology
  • Genetics
  • Molecular Biology

Cite this

Busch, D. B., Van Vuuren, H., De Wit, J., Collins, A., Zdzienicka, M. Z., Mitchell, D. L., ... Hoeijmakers, J. (1997). Phenotypic heterogeneity in nucleotide excision repair mutants of rodent complementation groups 1 and 4. Mutation Research - DNA Repair, 383(2), 91-106. https://doi.org/10.1016/S0921-8777(96)00048-1

Phenotypic heterogeneity in nucleotide excision repair mutants of rodent complementation groups 1 and 4. / Busch, David B.; Van Vuuren, Hanneke; De Wit, Jan; Collins, Andrew; Zdzienicka, Małgorzata Z.; Mitchell, David L.; Brookman, Kerry W.; Stefanini, Miria; Riboni, Roberta; Thompson, Larry H.; Albert, Roberta Bliss; Van Gool, Alain J.; Hoeijmakers, Jan.

In: Mutation Research - DNA Repair, Vol. 383, No. 2, 12.03.1997, p. 91-106.

Research output: Contribution to journalArticle

Busch, DB, Van Vuuren, H, De Wit, J, Collins, A, Zdzienicka, MZ, Mitchell, DL, Brookman, KW, Stefanini, M, Riboni, R, Thompson, LH, Albert, RB, Van Gool, AJ & Hoeijmakers, J 1997, 'Phenotypic heterogeneity in nucleotide excision repair mutants of rodent complementation groups 1 and 4', Mutation Research - DNA Repair, vol. 383, no. 2, pp. 91-106. https://doi.org/10.1016/S0921-8777(96)00048-1
Busch, David B. ; Van Vuuren, Hanneke ; De Wit, Jan ; Collins, Andrew ; Zdzienicka, Małgorzata Z. ; Mitchell, David L. ; Brookman, Kerry W. ; Stefanini, Miria ; Riboni, Roberta ; Thompson, Larry H. ; Albert, Roberta Bliss ; Van Gool, Alain J. ; Hoeijmakers, Jan. / Phenotypic heterogeneity in nucleotide excision repair mutants of rodent complementation groups 1 and 4. In: Mutation Research - DNA Repair. 1997 ; Vol. 383, No. 2. pp. 91-106.
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abstract = "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.",
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AU - Busch, David B.

AU - Van Vuuren, Hanneke

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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

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KW - Chinese hamster ovary (CHO)

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