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. 1997 Aug;17(8):4774–4781. doi: 10.1128/mcb.17.8.4774

Illegitimate recombination leading to allelic loss and unbalanced translocation in p53-mutated human lymphoblastoid cells.

M Honma 1, L S Zhang 1, M Hayashi 1, K Takeshita 1, Y Nakagawa 1, N Tanaka 1, T Sofuni 1
PMCID: PMC232329  PMID: 9234733

Abstract

Allelic loss and translocation are critical mutational events in human tumorigenesis. Allelic loss, which is usually identified as loss of heterozygosity (LOH), is frequently observed at tumor suppressor loci in various kinds of human tumors. It is generally thought to result from deletion or mitotic recombination between homologous chromosomes. In this report, we demonstrate that illegitimate (nonhomologous) recombination strongly contributes to the generation of allelic loss in p53-mutated cells. Spontaneous and X-ray-induced LOH mutations at the heterozygous thymidine kinase (tk) gene, which is located on the long arm of chromosome 17, from normal (TK6) and p53-mutated (WTK-1) human lymphoblastoid cells were cytogenetically analyzed by chromosome 17 painting. We observed unbalanced translocations in 53% of LOH mutants spontaneously arising from WTK-1 cells but none spontaneously arising from TK6 cells. We postulate that illegitimate recombination was occurring between nonhomologous chromosomes after DNA replication, leading to allelic loss and unbalanced translocations in p53-mutated WTK-1 cells. X-ray irradiation, which induces DNA double-strand breaks (DSBs), enhanced the generation of unbalanced translocation more efficiently in WTK-1 than in TK6 cells. This observation implicates the wild-type p53 protein in the regulation of homologous recombination and recombinational DNA repair of DSBs and suggests a possible mechanism by which loss of p53 function may cause genomic instability.

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Selected References

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  1. Agapova L. S., Ilyinskaya G. V., Turovets N. A., Ivanov A. V., Chumakov P. M., Kopnin B. P. Chromosome changes caused by alterations of p53 expression. Mutat Res. 1996 Jul 5;354(1):129–138. doi: 10.1016/0027-5107(96)00062-0. [DOI] [PubMed] [Google Scholar]
  2. Amundson S. A., Xia F., Wolfson K., Liber H. L. Different cytotoxic and mutagenic responses induced by X-rays in two human lymphoblastoid cell lines derived from a single donor. Mutat Res. 1993 Apr;286(2):233–241. doi: 10.1016/0027-5107(93)90188-l. [DOI] [PubMed] [Google Scholar]
  3. Bakalkin G., Yakovleva T., Selivanova G., Magnusson K. P., Szekely L., Kiseleva E., Klein G., Terenius L., Wiman K. G. p53 binds single-stranded DNA ends and catalyzes DNA renaturation and strand transfer. Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):413–417. doi: 10.1073/pnas.91.1.413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baumann P., Benson F. E., West S. C. Human Rad51 protein promotes ATP-dependent homologous pairing and strand transfer reactions in vitro. Cell. 1996 Nov 15;87(4):757–766. doi: 10.1016/s0092-8674(00)81394-x. [DOI] [PubMed] [Google Scholar]
  5. Bollag R. J., Waldman A. S., Liskay R. M. Homologous recombination in mammalian cells. Annu Rev Genet. 1989;23:199–225. doi: 10.1146/annurev.ge.23.120189.001215. [DOI] [PubMed] [Google Scholar]
  6. Bouffler S. D., Kemp C. J., Balmain A., Cox R. Spontaneous and ionizing radiation-induced chromosomal abnormalities in p53-deficient mice. Cancer Res. 1995 Sep 1;55(17):3883–3889. [PubMed] [Google Scholar]
  7. Chaganti R. S., Schonberg S., German J. A manyfold increase in sister chromatid exchanges in Bloom's syndrome lymphocytes. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4508–4512. doi: 10.1073/pnas.71.11.4508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cross S. M., Sanchez C. A., Morgan C. A., Schimke M. K., Ramel S., Idzerda R. L., Raskind W. H., Reid B. J. A p53-dependent mouse spindle checkpoint. Science. 1995 Mar 3;267(5202):1353–1356. doi: 10.1126/science.7871434. [DOI] [PubMed] [Google Scholar]
  9. Ellis N. A., Groden J., Ye T. Z., Straughen J., Lennon D. J., Ciocci S., Proytcheva M., German J. The Bloom's syndrome gene product is homologous to RecQ helicases. Cell. 1995 Nov 17;83(4):655–666. doi: 10.1016/0092-8674(95)90105-1. [DOI] [PubMed] [Google Scholar]
  10. Ellis N. A., Lennon D. J., Proytcheva M., Alhadeff B., Henderson E. E., German J. Somatic intragenic recombination within the mutated locus BLM can correct the high sister-chromatid exchange phenotype of Bloom syndrome cells. Am J Hum Genet. 1995 Nov;57(5):1019–1027. [PMC free article] [PubMed] [Google Scholar]
  11. Fearon E. R., Vogelstein B. A genetic model for colorectal tumorigenesis. Cell. 1990 Jun 1;61(5):759–767. doi: 10.1016/0092-8674(90)90186-i. [DOI] [PubMed] [Google Scholar]
  12. Fukasawa K., Choi T., Kuriyama R., Rulong S., Vande Woude G. F. Abnormal centrosome amplification in the absence of p53. Science. 1996 Mar 22;271(5256):1744–1747. doi: 10.1126/science.271.5256.1744. [DOI] [PubMed] [Google Scholar]
  13. GERMAN J., ARCHIBALD R., BLOOM D. CHROMOSOMAL BREAKAGE IN A RARE AND PROBABLY GENETICALLY DETERMINED SYNDROME OF MAN. Science. 1965 Apr 23;148(3669):506–507. doi: 10.1126/science.148.3669.506. [DOI] [PubMed] [Google Scholar]
  14. German J. Bloom syndrome: a mendelian prototype of somatic mutational disease. Medicine (Baltimore) 1993 Nov;72(6):393–406. [PubMed] [Google Scholar]
  15. Grosovsky A. J., Parks K. K., Giver C. R., Nelson S. L. Clonal analysis of delayed karyotypic abnormalities and gene mutations in radiation-induced genetic instability. Mol Cell Biol. 1996 Nov;16(11):6252–6262. doi: 10.1128/mcb.16.11.6252. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hartmann A., Speit G. Genotoxic effects of chemicals in the single cell gel (SCG) test with human blood cells in relation to the induction of sister-chromatid exchanges (SCE). Mutat Res. 1995 Jan;346(1):49–56. doi: 10.1016/0165-7992(95)90068-3. [DOI] [PubMed] [Google Scholar]
  17. Honma M., Hayashi M., Sofuni T. Cytotoxic and mutagenic responses to X-rays and chemical mutagens in normal and p53-mutated human lymphoblastoid cells. Mutat Res. 1997 Mar 4;374(1):89–98. doi: 10.1016/s0027-5107(96)00223-0. [DOI] [PubMed] [Google Scholar]
  18. Honma M., Little J. B. Recombinagenic activity of the phorbol ester 12-O-tetradecanoylphorbol-13-acetate in human lymphoblastoid cells. Carcinogenesis. 1995 Aug;16(8):1717–1722. doi: 10.1093/carcin/16.8.1717. [DOI] [PubMed] [Google Scholar]
  19. Kastan M. B., Onyekwere O., Sidransky D., Vogelstein B., Craig R. W. Participation of p53 protein in the cellular response to DNA damage. Cancer Res. 1991 Dec 1;51(23 Pt 1):6304–6311. [PubMed] [Google Scholar]
  20. Kern S. E., Kinzler K. W., Baker S. J., Nigro J. M., Rotter V., Levine A. J., Friedman P., Prives C., Vogelstein B. Mutant p53 proteins bind DNA abnormally in vitro. Oncogene. 1991 Jan;6(1):131–136. [PubMed] [Google Scholar]
  21. Kodama Y., Boreiko C. J., Skopek T. R., Recio L. Cytogenetic analysis of spontaneous and 2-cyanoethylene oxide-induced tk-/- mutants in TK6 human lymphoblastoid cultures. Environ Mol Mutagen. 1989;14(3):149–154. doi: 10.1002/em.2850140304. [DOI] [PubMed] [Google Scholar]
  22. Kuerbitz S. J., Plunkett B. S., Walsh W. V., Kastan M. B. Wild-type p53 is a cell cycle checkpoint determinant following irradiation. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7491–7495. doi: 10.1073/pnas.89.16.7491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lane D. P. Cancer. p53, guardian of the genome. Nature. 1992 Jul 2;358(6381):15–16. doi: 10.1038/358015a0. [DOI] [PubMed] [Google Scholar]
  24. Lasko D., Cavenee W., Nordenskjöld M. Loss of constitutional heterozygosity in human cancer. Annu Rev Genet. 1991;25:281–314. doi: 10.1146/annurev.ge.25.120191.001433. [DOI] [PubMed] [Google Scholar]
  25. Lee J. M., Bernstein A. p53 mutations increase resistance to ionizing radiation. Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5742–5746. doi: 10.1073/pnas.90.12.5742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lim D. S., Hasty P. A mutation in mouse rad51 results in an early embryonic lethal that is suppressed by a mutation in p53. Mol Cell Biol. 1996 Dec;16(12):7133–7143. doi: 10.1128/mcb.16.12.7133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Little J. B., Nagasawa H., Keng P. C., Yu Y., Li C. Y. Absence of radiation-induced G1 arrest in two closely related human lymphoblast cell lines that differ in p53 status. J Biol Chem. 1995 May 12;270(19):11033–11036. doi: 10.1074/jbc.270.19.11033. [DOI] [PubMed] [Google Scholar]
  28. Livingstone L. R., White A., Sprouse J., Livanos E., Jacks T., Tlsty T. D. Altered cell cycle arrest and gene amplification potential accompany loss of wild-type p53. Cell. 1992 Sep 18;70(6):923–935. doi: 10.1016/0092-8674(92)90243-6. [DOI] [PubMed] [Google Scholar]
  29. Loeb L. A., Christians F. C. Multiple mutations in human cancers. Mutat Res. 1996 Feb 19;350(1):279–286. doi: 10.1016/0027-5107(95)00117-4. [DOI] [PubMed] [Google Scholar]
  30. Lu X., Lane D. P. Differential induction of transcriptionally active p53 following UV or ionizing radiation: defects in chromosome instability syndromes? Cell. 1993 Nov 19;75(4):765–778. doi: 10.1016/0092-8674(93)90496-d. [DOI] [PubMed] [Google Scholar]
  31. Morgan W. F., Day J. P., Kaplan M. I., McGhee E. M., Limoli C. L. Genomic instability induced by ionizing radiation. Radiat Res. 1996 Sep;146(3):247–258. [PubMed] [Google Scholar]
  32. Mummenbrauer T., Janus F., Müller B., Wiesmüller L., Deppert W., Grosse F. p53 Protein exhibits 3'-to-5' exonuclease activity. Cell. 1996 Jun 28;85(7):1089–1099. doi: 10.1016/s0092-8674(00)81309-4. [DOI] [PubMed] [Google Scholar]
  33. Nowell P. C. The clonal evolution of tumor cell populations. Science. 1976 Oct 1;194(4260):23–28. doi: 10.1126/science.959840. [DOI] [PubMed] [Google Scholar]
  34. Oberosler P., Hloch P., Ramsperger U., Stahl H. p53-catalyzed annealing of complementary single-stranded nucleic acids. EMBO J. 1993 Jun;12(6):2389–2396. doi: 10.1002/j.1460-2075.1993.tb05893.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Roth D. B., Lindahl T., Gellert M. Repair and recombination. How to make ends meet. Curr Biol. 1995 May 1;5(5):496–499. doi: 10.1016/s0960-9822(95)00101-1. [DOI] [PubMed] [Google Scholar]
  36. Sands A. T., Suraokar M. B., Sanchez A., Marth J. E., Donehower L. A., Bradley A. p53 deficiency does not affect the accumulation of point mutations in a transgene target. Proc Natl Acad Sci U S A. 1995 Aug 29;92(18):8517–8521. doi: 10.1073/pnas.92.18.8517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Selivanova G., Wiman K. G. p53: a cell cycle regulator activated by DNA damage. Adv Cancer Res. 1995;66:143–180. doi: 10.1016/s0065-230x(08)60253-5. [DOI] [PubMed] [Google Scholar]
  38. Sengstag C. The role of mitotic recombination in carcinogenesis. Crit Rev Toxicol. 1994;24(4):323–353. doi: 10.3109/10408449409017922. [DOI] [PubMed] [Google Scholar]
  39. Shinohara A., Ogawa H., Matsuda Y., Ushio N., Ikeo K., Ogawa T. Cloning of human, mouse and fission yeast recombination genes homologous to RAD51 and recA. Nat Genet. 1993 Jul;4(3):239–243. doi: 10.1038/ng0793-239. [DOI] [PubMed] [Google Scholar]
  40. Stahl F. W. Roles of double-strand breaks in generalized genetic recombination. Prog Nucleic Acid Res Mol Biol. 1986;33:169–194. doi: 10.1016/s0079-6603(08)60023-9. [DOI] [PubMed] [Google Scholar]
  41. Stürzbecher H. W., Donzelmann B., Henning W., Knippschild U., Buchhop S. p53 is linked directly to homologous recombination processes via RAD51/RecA protein interaction. EMBO J. 1996 Apr 15;15(8):1992–2002. [PMC free article] [PubMed] [Google Scholar]
  42. Suwa A., Hirakata M., Takeda Y., Jesch S. A., Mimori T., Hardin J. A. DNA-dependent protein kinase (Ku protein-p350 complex) assembles on double-stranded DNA. Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):6904–6908. doi: 10.1073/pnas.91.15.6904. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Symonds H., Krall L., Remington L., Saenz-Robles M., Lowe S., Jacks T., Van Dyke T. p53-dependent apoptosis suppresses tumor growth and progression in vivo. Cell. 1994 Aug 26;78(4):703–711. doi: 10.1016/0092-8674(94)90534-7. [DOI] [PubMed] [Google Scholar]
  44. Tachibana A., Tatsumi K., Masui T., Kato T. Large deletions at the HPRT locus associated with the mutator phenotype in a Bloom's syndrome lymphoblastoid cell line. Mol Carcinog. 1996 Sep;17(1):41–47. doi: 10.1002/(SICI)1098-2744(199609)17:1<41::AID-MC6>3.0.CO;2-N. [DOI] [PubMed] [Google Scholar]
  45. Tlsty T. D., Briot A., Gualberto A., Hall I., Hess S., Hixon M., Kuppuswamy D., Romanov S., Sage M., White A. Genomic instability and cancer. Mutat Res. 1995 Jul;337(1):1–7. doi: 10.1016/0921-8777(95)00016-d. [DOI] [PubMed] [Google Scholar]
  46. Xia F., Amundson S. A., Nickoloff J. A., Liber H. L. Different capacities for recombination in closely related human lymphoblastoid cell lines with different mutational responses to X-irradiation. Mol Cell Biol. 1994 Sep;14(9):5850–5857. doi: 10.1128/mcb.14.9.5850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Xia F., Wang X., Wang Y. H., Tsang N. M., Yandell D. W., Kelsey K. T., Liber H. L. Altered p53 status correlates with differences in sensitivity to radiation-induced mutation and apoptosis in two closely related human lymphoblast lines. Cancer Res. 1995 Jan 1;55(1):12–15. [PubMed] [Google Scholar]
  48. Yin Y., Tainsky M. A., Bischoff F. Z., Strong L. C., Wahl G. M. Wild-type p53 restores cell cycle control and inhibits gene amplification in cells with mutant p53 alleles. Cell. 1992 Sep 18;70(6):937–948. doi: 10.1016/0092-8674(92)90244-7. [DOI] [PubMed] [Google Scholar]
  49. Zhang L. S., Honma M., Matsuoka A., Suzuki T., Sofuni T., Hayashi M. Chromosome painting analysis of spontaneous and methyl methanesulfonate-induced trifluorothymidine-resistant L5178Y cell colonies. Mutat Res. 1996 Oct 1;370(3-4):181–190. doi: 10.1016/s0165-1218(96)00069-9. [DOI] [PubMed] [Google Scholar]

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