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. 1986 Jun;83(12):4379–4383. doi: 10.1073/pnas.83.12.4379

Locus specificity in the mutability of L5178Y mouse lymphoma cells: the role of multilocus lesions.

H H Evans, J Mencl, M F Horng, M Ricanati, C Sanchez, J Hozier
PMCID: PMC323736  PMID: 3012566

Abstract

Mouse L5178Y strain LY-S and its parental strain LY-R differ in their comparative sensitivities to the cytotoxic effects of various mutagenic agents--i.e., strain LY-S has been found to be more sensitive, less sensitive, or similarly sensitive to individual agents in comparison to strain LY-R. Nevertheless, strain LY-S has been found to be uniformly less mutable than strain LY-R at the hypoxanthine (guanine) phosphoribosyltransferase (Hprt) locus following treatment with x-radiation, UV radiation, or alkylating agents. In the present work we have isolated subclones of strains LY-R and LY-S that are heterozygous at the thymidine kinase (Tk) locus (chromosome 11). We have found that a heterozygous LY-S Tk+/Tk- strain shows as high or higher mutability at the Tk locus than do heterozygous LY-R strains following treatment with x-radiation, UV radiation, or ethyl methanesulfonate. Mutability of all heterozygous strains at the Tk locus is much higher than at the Hprt locus following treatment with these mutagenic agents, with the exception of one heterozygous LY-R strain that possesses only one chromosome 11 and that is poorly mutable at the Tk locus by x-radiation. On the basis of these results, we have suggested that because of a repair deficiency, multilocus lesions are formed in the DNA of LY-S strains following treatment with radiation or alkylating agents; multilocus lesions lead to poor recovery of viable mutants when the target locus is closely linked to essential genes and situated on a hemizygous chromosomal region (e.g., the Hprt locus on the X chromosome or the Tk locus in strains monosomic for chromosome 11); and x-radiation is a relatively poor mutagen at loci situated on hemizygous chromosomal regions, in repair-efficient and repair-deficient cells, because of its tendency to form multilocus lesions.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ALEXANDER P. Mouse lymphoma cells with different radiosensitivities. Nature. 1961 Nov 11;192:572–573. doi: 10.1038/192572a0. [DOI] [PubMed] [Google Scholar]
  2. Abrahamson S., Wolff S. Re-analysis of radiation-induced specific locus mutations in the mouse. Nature. 1976 Dec 23;264(5588):715–719. doi: 10.1038/264715a0. [DOI] [PubMed] [Google Scholar]
  3. Arlett C. F., Turnbull D., Harcourt S. A., Lehmann A. R., Colella C. M. A comparison of the 8-azaguanine and ouabain-resistance systems for the selection of induced mutant Chinese hamster cells. Mutat Res. 1975 Dec;33(2-3):261–278. doi: 10.1016/0027-5107(75)90202-x. [DOI] [PubMed] [Google Scholar]
  4. BREITMAN T. R. The feedback inhibition of thymidine kinase. Biochim Biophys Acta. 1963 Jan 8;67:153–155. doi: 10.1016/0006-3002(63)91807-9. [DOI] [PubMed] [Google Scholar]
  5. Beer J. Z., Budzicka E., Niepokojczycka E., Rosiek O., Szumiel I., Walicka M. Loss of tumorigenicity with simultaneous changes in radiosensitivity and photosensitivity during in vitro growth of L5178Y murine lymphoma cells. Cancer Res. 1983 Oct;43(10):4736–4742. [PubMed] [Google Scholar]
  6. Beer J. Z., Jacobson E. D., Evans H. H., Szumiel I. X-ray and UV mutagenesis in two L5178Y cell strains differing in tumorigenicity, radiosensitivity, and DNA repair. Br J Cancer Suppl. 1984;6:107–111. [PMC free article] [PubMed] [Google Scholar]
  7. Bocian E., Bouzyk E., Rosiek O., Ziemba-Zótowska B. Chromosomal aberrations induced by x-rays in two mouse lymphoma (L5178Y) sublines of different radiosensitivity. Int J Radiat Biol Relat Stud Phys Chem Med. 1982 Sep;42(3):347–351. doi: 10.1080/09553008214551261. [DOI] [PubMed] [Google Scholar]
  8. Clive D., Johnson K. O., Spector J. F., Batson A. G., Brown M. M. Validation and characterization of the L5178Y/TK+/- mouse lymphoma mutagen assay system. Mutat Res. 1979 Jan;59(1):61–108. doi: 10.1016/0027-5107(79)90195-7. [DOI] [PubMed] [Google Scholar]
  9. Clive D., Spector J. F. Laboratory procedure for assessing specific locus mutations at the TK locus in cultured L5178Y mouse lymphoma cells. Mutat Res. 1975 Feb;31(1):17–29. doi: 10.1016/0165-1161(75)90059-x. [DOI] [PubMed] [Google Scholar]
  10. Clive D., Voytek P. Evidence for chemically-induced structural gene mutations at the thymidine kinase locus in cultured L5178Y mouse lymphoma cells. Mutat Res. 1977 Aug;44(2):269–278. doi: 10.1016/0027-5107(77)90084-7. [DOI] [PubMed] [Google Scholar]
  11. Cox R., Masson W. K. Do radiation-induced thioguanine-resistant mutants of cultured mammalian cells arise by HGPRT gene mutation or X-chromosome rearrangement? Nature. 1978 Dec 7;276(5688):629–630. doi: 10.1038/276629a0. [DOI] [PubMed] [Google Scholar]
  12. Cox R., Thacker J., Goodhead D. T., Munson R. J. Mutation and inactivation of mammalian cells by various ionising radiations. Nature. 1977 Jun 2;267(5610):425–427. doi: 10.1038/267425a0. [DOI] [PubMed] [Google Scholar]
  13. Evans H. H., Horng M. F., Beer J. Z. Lethal and mutagenic effects of radiation and alkylating agents on two strains of mouse L5178Y cells. Mutat Res. 1986 Jun;161(1):91–97. doi: 10.1016/0027-5107(86)90103-x. [DOI] [PubMed] [Google Scholar]
  14. Evans H. H., Horng M. F., Mencl J., Glazier K. G., Beer J. Z. The influence of dose rate on the lethal and mutagenic effects of X-rays in proliferating L5178Y cells differing in radiation sensitivity. Int J Radiat Biol Relat Stud Phys Chem Med. 1985 May;47(5):553–562. doi: 10.1080/09553008514550781. [DOI] [PubMed] [Google Scholar]
  15. Graf L. H., Jr, Chasin L. A. Direct demonstration of genetic alterations at the dihydrofolate reductase locus after gamma irradiation. Mol Cell Biol. 1982 Jan;2(1):93–96. doi: 10.1128/mcb.2.1.93. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hozier J., Sawyer J., Clive D., Moore M. M. Chromosome 11 aberrations in small colony L5178Y TK-/- mutants early in their clonal history. Mutat Res. 1985 Oct;147(5):237–242. doi: 10.1016/0165-1161(85)90064-0. [DOI] [PubMed] [Google Scholar]
  17. Hozier J., Sawyer J., Moore M., Howard B., Clive D. Cytogenetic analysis of the L5178Y/TK+/- leads to TK-/- mouse lymphoma mutagenesis assay system. Mutat Res. 1981 Nov;84(1):169–181. doi: 10.1016/0027-5107(81)90060-9. [DOI] [PubMed] [Google Scholar]
  18. Jacobson E. D., Krell K., Olempska-Beer Z., Beer J. Z. UV-induced mutagenesis at the hypoxanthine-guanine phosphoribosyl transferase locus in two L5178Y mouse lymphoma cell strains with different UV sensitivities. Mutat Res. 1984 Nov;129(2):259–267. doi: 10.1016/0027-5107(84)90159-3. [DOI] [PubMed] [Google Scholar]
  19. Kavathas P., Bach F. H., DeMars R. Gamma ray-induced loss of expression of HLA and glyoxalase I alleles in lymphoblastoid cells. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4251–4255. doi: 10.1073/pnas.77.7.4251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kozak C. A., Ruddle F. H. Assignment of the genes for thymidine kinase and galactokinase to Mus musculus chromosome 11 and the preferential segregation of this chromosome in Chinese hamster/mouse somatic cell hybrids. Somatic Cell Genet. 1977 Mar;3(2):121–133. doi: 10.1007/BF01551809. [DOI] [PubMed] [Google Scholar]
  21. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  22. Lever J. E., Seegmiller J. E. Ouabain-resistant human lymphoblastoid lines altered in the (Na+ + K+)-dependent ATPase membrane transport system. J Cell Physiol. 1976 Jul;88(3):343–352. doi: 10.1002/jcp.1040880310. [DOI] [PubMed] [Google Scholar]
  23. Mankovitz R., Buchwald M., Baker R. M. Isolation of ouabain-resistant human diploid fibroblasts. Cell. 1974 Nov;3(3):221–226. doi: 10.1016/0092-8674(74)90135-4. [DOI] [PubMed] [Google Scholar]
  24. McBreen P., Orkwiszewski K. G., Chern C. J., Mellman W. J., Croce C. M. Synteny of the genes for thymidine kinase and galactokinase in the mouse and their assignment to mouse chromosome 11. Cytogenet Cell Genet. 1977;19(1):7–13. doi: 10.1159/000130789. [DOI] [PubMed] [Google Scholar]
  25. Moore M. M., Clive D., Hozier J. C., Howard B. E., Batson A. G., Turner N. T., Sawyer J. Analysis of trifluorothymidine-resistant (TFTr) mutants of L5178Y/TK+/- mouse lymphoma cells. Mutat Res. 1985 Aug;151(1):161–174. doi: 10.1016/0027-5107(85)90194-0. [DOI] [PubMed] [Google Scholar]
  26. Moore M. M., Clive D. The quantitation of TK-/- and HGPRT- mutants of L5178Y/TK+/- mouse lymphoma cells at varying times post-treatment. Environ Mutagen. 1982;4(4):499–519. doi: 10.1002/em.2860040409. [DOI] [PubMed] [Google Scholar]
  27. Schneider E. L., Stanbridge E. J., Epstein C. J. Incorporation of 3H-uridine and 3H-uracil into RNA: a simple technique for the detection of mycoplasma contamination of cultured cells. Exp Cell Res. 1974 Mar 15;84(1):311–318. doi: 10.1016/0014-4827(74)90411-x. [DOI] [PubMed] [Google Scholar]
  28. Thacker J., Stephens M. A., Stretch A. Mutation to ouabain-resistance in Chinese hamster cells: induction by ethyl methanesulphonate and lack of induction by ionising radiation. Mutat Res. 1978 Aug;51(2):255–270. doi: 10.1016/s0027-5107(78)80021-9. [DOI] [PubMed] [Google Scholar]
  29. WEBBER B. B., DESERRES F. J. INDUCTION KINETICS AND GENETIC ANALYSIS OF X-RAY-INDUCED MUTATIONS IN THE AD-3 REGION OF NEUROSPORA CRASSA. Proc Natl Acad Sci U S A. 1965 Feb;53:430–437. doi: 10.1073/pnas.53.2.430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Waldren C., Jones C., Puck T. T. Measurement of mutagenesis in mammalian cells. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1358–1362. doi: 10.1073/pnas.76.3.1358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Yau T. M., Kim S. C., Gregg E. C., Nygaard O. F. Inverse X-irradiation split-dose effect in a murine lymphoma cell line. Int J Radiat Biol Relat Stud Phys Chem Med. 1979 Jun;35(6):577–581. doi: 10.1080/09553007914550691. [DOI] [PubMed] [Google Scholar]
  32. de Serres F. J., Inoue H., Schüpbach M. E. Mutagenesis at the ad-3A and ad-3B loci in haploid UV-sensitive strains of Neurospora crassa. VI. Genetic characterization of ad-3 mutants provides evidence for qualitative differences in the spectrum of genetic alterations between wild-type and nucleotide excision-repair-deficient strains. Mutat Res. 1983 Mar;108(1-3):93–108. doi: 10.1016/0027-5107(83)90112-4. [DOI] [PubMed] [Google Scholar]

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