Carcinogens can induce homologous recombination between duplicated chromosomal sequences in mouse L cells

Y. Wang, V. M. Maher, Robert (Mike) Liskay, J. J. McCormick

Research output: Contribution to journalArticle

98 Citations (Scopus)

Abstract

The ability of a series of DNA-damaging agents to induce homologous intrachromosomal recombination between duplicated genes in the chromosome of mouse cells was investigated. The target cells were the thymidine kinase-deficient mouse L-cell strain 333M, which contains a single integrated copy of a plasmid with two herpes simplex virus thymidine kinase (Htk) genes, each containing an 8-base-pair XhoI linker inserted at a unique site. Expression of a functional Htk enzyme requires a productive recombinational event between the two nonfunctional genes. The spontaneous rate of recombination in this strain is 3 per 106 cells per generation. The agents tested represent physical carcinogens (UV and ionizing radiation), a simple alkylating agent (N-methyl-N'-nitro-N-nitrosoguanidine), an alkylating cross-linking agent (mitomycin C), and a reactive metabolite of a polycyclic aromatic hydrocarbon {(±)-7β,8α-dihydroxy-9α,10α-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene [BPDE]}. The background frequency of tk+ recombinants in the untreated population averaged 18 x 10-6 ± 5 x 10-6. Ionizing radiation had little or no effect on recombination; exposure to mitomycin C, N-methyl-N'-nitro-N-nitrosoguanidine, BPDE, or UV, at doses that lowered the survival to between 90 and 10% of the control, caused a dose-dependent increase in frequency of recombinants, reaching 50 x 10-6 to 100 x 10-6. No tk+ cells could be generated with a control cell line that contained only one mutant copy of the Htk gene. Molecular hybridization analysis showed that 85 to 90% of the tk+ recombinants retained the Htk gene duplication, consistent with nonreciprocal transfer of wild-type genetic information, gene conversion. In the rest, only a single copy of the Htk gene remained, reflecting a single reciprocal exchange within a chromatid or a single unequal exchange between sister chromatids. Each recombinant tested contained an XhoI-resistant (wild-type) Htk gene.

Original languageEnglish (US)
Pages (from-to)196-202
Number of pages7
JournalMolecular and Cellular Biology
Volume8
Issue number1
StatePublished - 1988
Externally publishedYes

Fingerprint

Homologous Recombination
Carcinogens
7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide
Genes
Methylnitronitrosoguanidine
Thymidine Kinase
Mitomycin
Ionizing Radiation
Genetic Recombination
Gene Conversion
Sister Chromatid Exchange
Chromatids
Gene Duplication
Alkylating Agents
Polycyclic Aromatic Hydrocarbons
Simplexvirus
Base Pairing
Plasmids
Chromosomes
Cell Line

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

Carcinogens can induce homologous recombination between duplicated chromosomal sequences in mouse L cells. / Wang, Y.; Maher, V. M.; Liskay, Robert (Mike); McCormick, J. J.

In: Molecular and Cellular Biology, Vol. 8, No. 1, 1988, p. 196-202.

Research output: Contribution to journalArticle

Wang, Y. ; Maher, V. M. ; Liskay, Robert (Mike) ; McCormick, J. J. / Carcinogens can induce homologous recombination between duplicated chromosomal sequences in mouse L cells. In: Molecular and Cellular Biology. 1988 ; Vol. 8, No. 1. pp. 196-202.
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abstract = "The ability of a series of DNA-damaging agents to induce homologous intrachromosomal recombination between duplicated genes in the chromosome of mouse cells was investigated. The target cells were the thymidine kinase-deficient mouse L-cell strain 333M, which contains a single integrated copy of a plasmid with two herpes simplex virus thymidine kinase (Htk) genes, each containing an 8-base-pair XhoI linker inserted at a unique site. Expression of a functional Htk enzyme requires a productive recombinational event between the two nonfunctional genes. The spontaneous rate of recombination in this strain is 3 per 106 cells per generation. The agents tested represent physical carcinogens (UV and ionizing radiation), a simple alkylating agent (N-methyl-N'-nitro-N-nitrosoguanidine), an alkylating cross-linking agent (mitomycin C), and a reactive metabolite of a polycyclic aromatic hydrocarbon {(±)-7β,8α-dihydroxy-9α,10α-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene [BPDE]}. The background frequency of tk+ recombinants in the untreated population averaged 18 x 10-6 ± 5 x 10-6. Ionizing radiation had little or no effect on recombination; exposure to mitomycin C, N-methyl-N'-nitro-N-nitrosoguanidine, BPDE, or UV, at doses that lowered the survival to between 90 and 10{\%} of the control, caused a dose-dependent increase in frequency of recombinants, reaching 50 x 10-6 to 100 x 10-6. No tk+ cells could be generated with a control cell line that contained only one mutant copy of the Htk gene. Molecular hybridization analysis showed that 85 to 90{\%} of the tk+ recombinants retained the Htk gene duplication, consistent with nonreciprocal transfer of wild-type genetic information, gene conversion. In the rest, only a single copy of the Htk gene remained, reflecting a single reciprocal exchange within a chromatid or a single unequal exchange between sister chromatids. Each recombinant tested contained an XhoI-resistant (wild-type) Htk gene.",
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N2 - The ability of a series of DNA-damaging agents to induce homologous intrachromosomal recombination between duplicated genes in the chromosome of mouse cells was investigated. The target cells were the thymidine kinase-deficient mouse L-cell strain 333M, which contains a single integrated copy of a plasmid with two herpes simplex virus thymidine kinase (Htk) genes, each containing an 8-base-pair XhoI linker inserted at a unique site. Expression of a functional Htk enzyme requires a productive recombinational event between the two nonfunctional genes. The spontaneous rate of recombination in this strain is 3 per 106 cells per generation. The agents tested represent physical carcinogens (UV and ionizing radiation), a simple alkylating agent (N-methyl-N'-nitro-N-nitrosoguanidine), an alkylating cross-linking agent (mitomycin C), and a reactive metabolite of a polycyclic aromatic hydrocarbon {(±)-7β,8α-dihydroxy-9α,10α-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene [BPDE]}. The background frequency of tk+ recombinants in the untreated population averaged 18 x 10-6 ± 5 x 10-6. Ionizing radiation had little or no effect on recombination; exposure to mitomycin C, N-methyl-N'-nitro-N-nitrosoguanidine, BPDE, or UV, at doses that lowered the survival to between 90 and 10% of the control, caused a dose-dependent increase in frequency of recombinants, reaching 50 x 10-6 to 100 x 10-6. No tk+ cells could be generated with a control cell line that contained only one mutant copy of the Htk gene. Molecular hybridization analysis showed that 85 to 90% of the tk+ recombinants retained the Htk gene duplication, consistent with nonreciprocal transfer of wild-type genetic information, gene conversion. In the rest, only a single copy of the Htk gene remained, reflecting a single reciprocal exchange within a chromatid or a single unequal exchange between sister chromatids. Each recombinant tested contained an XhoI-resistant (wild-type) Htk gene.

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