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Mol Cell Biol. Oct 1997; 17(10): 6087–6096.
PMCID: PMC232458

hMre11 and hRad50 nuclear foci are induced during the normal cellular response to DNA double-strand breaks.

Abstract

We previously identified a conserved multiprotein complex that includes hMre11 and hRad50. In this study, we used immunofluorescence to investigate the role of this complex in DNA double-strand break (DSB) repair. hMre11 and hRad50 form discrete nuclear foci in response to treatment with DSB-inducing agents but not in response to UV irradiation. hMre11 and hRad50 foci colocalize after treatment with ionizing radiation and are distinct from those of the DSB repair protein, hRad51. Our data indicate that an irradiated cell is competent to form either hMre11-hRad50 foci or hRad51 foci, but not both. The multiplicity of hMre11 and hRad50 foci is much higher in the DSB repair-deficient cell line 180BR than in repair-proficient cells. hMre11-hRad50 focus formation is markedly reduced in cells derived from ataxia-telangiectasia patients, whereas hRad51 focus formation is markedly increased. These experiments support genetic evidence from Saccharomyces cerevisiae indicating that Mre11-Rad50 have roles distinct from that of Rad51 in DSB repair. Further, these data indicate that hMre11-hRad50 foci form in response to DNA DSBs and are dependent upon a DNA damage-induced signaling pathway.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Ajimura M, Leem SH, Ogawa H. Identification of new genes required for meiotic recombination in Saccharomyces cerevisiae. Genetics. 1993 Jan;133(1):51–66. [PMC free article] [PubMed]
  • Allalunis-Turner MJ, Barron GM, Day RS, 3rd, Dobler KD, Mirzayans R. Isolation of two cell lines from a human malignant glioma specimen differing in sensitivity to radiation and chemotherapeutic drugs. Radiat Res. 1993 Jun;134(3):349–354. [PubMed]
  • Badie C, Iliakis G, Foray N, Alsbeih G, Cedervall B, Chavaudra N, Pantelias G, Arlett C, Malaise EP. Induction and rejoining of DNA double-strand breaks and interphase chromosome breaks after exposure to X rays in one normal and two hypersensitive human fibroblast cell lines. Radiat Res. 1995 Oct;144(1):26–35. [PubMed]
  • Badie C, Iliakis G, Foray N, Alsbeih G, Pantellias GE, Okayasu R, Cheong N, Russell NS, Begg AC, Arlett CF, et al. Defective repair of DNA double-strand breaks and chromosome damage in fibroblasts from a radiosensitive leukemia patient. Cancer Res. 1995 Mar 15;55(6):1232–1234. [PubMed]
  • Bai Y, Symington LS. A Rad52 homolog is required for RAD51-independent mitotic recombination in Saccharomyces cerevisiae. Genes Dev. 1996 Aug 15;10(16):2025–2037. [PubMed]
  • Baker SM, Plug AW, Prolla TA, Bronner CE, Harris AC, Yao X, Christie DM, Monell C, Arnheim N, Bradley A, et al. Involvement of mouse Mlh1 in DNA mismatch repair and meiotic crossing over. Nat Genet. 1996 Jul;13(3):336–342. [PubMed]
  • Bendixen C, Sunjevaric I, Bauchwitz R, Rothstein R. Identification of a mouse homologue of the Saccharomyces cerevisiae recombination and repair gene, RAD52. Genomics. 1994 Sep 1;23(1):300–303. [PubMed]
  • Bishop DK. RecA homologs Dmc1 and Rad51 interact to form multiple nuclear complexes prior to meiotic chromosome synapsis. Cell. 1994 Dec 16;79(6):1081–1092. [PubMed]
  • Bollag RJ, Waldman AS, Liskay RM. Homologous recombination in mammalian cells. Annu Rev Genet. 1989;23:199–225. [PubMed]
  • Brown JM, Evans JW, Kovacs MS. Mechanism of chromosome exchange formation in human fibroblasts: insights from "chromosome painting". Environ Mol Mutagen. 1993;22(4):218–224. [PubMed]
  • Chen GL, Yang L, Rowe TC, Halligan BD, Tewey KM, Liu LF. Nonintercalative antitumor drugs interfere with the breakage-reunion reaction of mammalian DNA topoisomerase II. J Biol Chem. 1984 Nov 10;259(21):13560–13566. [PubMed]
  • Chow KC, Ross WE. Topoisomerase-specific drug sensitivity in relation to cell cycle progression. Mol Cell Biol. 1987 Sep;7(9):3119–3123. [PMC free article] [PubMed]
  • Cornforth MN, Bedford JS. On the nature of a defect in cells from individuals with ataxia-telangiectasia. Science. 1985 Mar 29;227(4694):1589–1591. [PubMed]
  • Cornforth MN, Bedford JS. X-ray--induced breakage and rejoining of human interphase chromosomes. Science. 1983 Dec 9;222(4628):1141–1143. [PubMed]
  • Dolganov GM, Maser RS, Novikov A, Tosto L, Chong S, Bressan DA, Petrini JH. Human Rad50 is physically associated with human Mre11: identification of a conserved multiprotein complex implicated in recombinational DNA repair. Mol Cell Biol. 1996 Sep;16(9):4832–4841. [PMC free article] [PubMed]
  • Dyck JA, Maul GG, Miller WH, Jr, Chen JD, Kakizuka A, Evans RM. A novel macromolecular structure is a target of the promyelocyte-retinoic acid receptor oncoprotein. Cell. 1994 Jan 28;76(2):333–343. [PubMed]
  • Flygare J, Benson F, Hellgren D. Expression of the human RAD51 gene during the cell cycle in primary human peripheral blood lymphocytes. Biochim Biophys Acta. 1996 Jul 24;1312(3):231–236. [PubMed]
  • Ford MD, Martin L, Lavin MF. The effects of ionizing radiation on cell cycle progression in ataxia telangiectasia. Mutat Res. 1984 Jan;125(1):115–122. [PubMed]
  • Fu XD, Maniatis T. Factor required for mammalian spliceosome assembly is localized to discrete regions in the nucleus. Nature. 1990 Feb 1;343(6257):437–441. [PubMed]
  • Game JC. DNA double-strand breaks and the RAD50-RAD57 genes in Saccharomyces. Semin Cancer Biol. 1993 Apr;4(2):73–83. [PubMed]
  • Giaccia A, Weinstein R, Hu J, Stamato TD. Cell cycle-dependent repair of double-strand DNA breaks in a gamma-ray-sensitive Chinese hamster cell. Somat Cell Mol Genet. 1985 Sep;11(5):485–491. [PubMed]
  • Haaf T, Golub EI, Reddy G, Radding CM, Ward DC. Nuclear foci of mammalian Rad51 recombination protein in somatic cells after DNA damage and its localization in synaptonemal complexes. Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):2298–2302. [PMC free article] [PubMed]
  • Haber JE. Exploring the pathways of homologous recombination. Curr Opin Cell Biol. 1992 Jun;4(3):401–412. [PubMed]
  • Heck MM, Hittelman WN, Earnshaw WC. Differential expression of DNA topoisomerases I and II during the eukaryotic cell cycle. Proc Natl Acad Sci U S A. 1988 Feb;85(4):1086–1090. [PMC free article] [PubMed]
  • Houldsworth J, Lavin MF. Effect of ionizing radiation on DNA synthesis in ataxia telangiectasia cells. Nucleic Acids Res. 1980 Aug 25;8(16):3709–3720. [PMC free article] [PubMed]
  • Hozák P, Hassan AB, Jackson DA, Cook PR. Visualization of replication factories attached to nucleoskeleton. Cell. 1993 Apr 23;73(2):361–373. [PubMed]
  • Hsiang YH, Wu HY, Liu LF. Proliferation-dependent regulation of DNA topoisomerase II in cultured human cells. Cancer Res. 1988 Jun 1;48(11):3230–3235. [PubMed]
  • Iliakis G. The role of DNA double strand breaks in ionizing radiation-induced killing of eukaryotic cells. Bioessays. 1991 Dec;13(12):641–648. [PubMed]
  • Ivanov EL, Korolev VG, Fabre F. XRS2, a DNA repair gene of Saccharomyces cerevisiae, is needed for meiotic recombination. Genetics. 1992 Nov;132(3):651–664. [PMC free article] [PubMed]
  • Jackson SP. DNA damage detection by DNA dependent protein kinase and related enzymes. Cancer Surv. 1996;28:261–279. [PubMed]
  • Jackson SP, Jeggo PA. DNA double-strand break repair and V(D)J recombination: involvement of DNA-PK. Trends Biochem Sci. 1995 Oct;20(10):412–415. [PubMed]
  • Jung M, Kondratyev A, Lee SA, Dimtchev A, Dritschilo A. ATM gene product phosphorylates I kappa B-alpha. Cancer Res. 1997 Jan 1;57(1):24–27. [PubMed]
  • Kanaar R, Troelstra C, Swagemakers SM, Essers J, Smit B, Franssen JH, Pastink A, Bezzubova OY, Buerstedde JM, Clever B, et al. Human and mouse homologs of the Saccharomyces cerevisiae RAD54 DNA repair gene: evidence for functional conservation. Curr Biol. 1996 Jul 1;6(7):828–838. [PubMed]
  • Kastan MB, Zhan Q, el-Deiry WS, Carrier F, Jacks T, Walsh WV, Plunkett BS, Vogelstein B, Fornace AJ., Jr A mammalian cell cycle checkpoint pathway utilizing p53 and GADD45 is defective in ataxia-telangiectasia. Cell. 1992 Nov 13;71(4):587–597. [PubMed]
  • Keegan KS, Holtzman DA, Plug AW, Christenson ER, Brainerd EE, Flaggs G, Bentley NJ, Taylor EM, Meyn MS, Moss SB, et al. The Atr and Atm protein kinases associate with different sites along meiotically pairing chromosomes. Genes Dev. 1996 Oct 1;10(19):2423–2437. [PubMed]
  • Khanna KK, Beamish H, Yan J, Hobson K, Williams R, Dunn I, Lavin MF. Nature of G1/S cell cycle checkpoint defect in ataxia-telangiectasia. Oncogene. 1995 Aug 17;11(4):609–618. [PubMed]
  • Lavin MF, Khanna KK, Beamish H, Teale B, Hobson K, Watters D. Defect in radiation signal transduction in ataxia-telangiectasia. Int J Radiat Biol. 1994 Dec;66(6 Suppl):S151–S156. [PubMed]
  • Lees-Miller SP, Godbout R, Chan DW, Weinfeld M, Day RS, 3rd, Barron GM, Allalunis-Turner J. Absence of p350 subunit of DNA-activated protein kinase from a radiosensitive human cell line. Science. 1995 Feb 24;267(5201):1183–1185. [PubMed]
  • Levine AJ. Tumor suppressor genes. Bioessays. 1990 Feb;12(2):60–66. [PubMed]
  • Li MJ, Peakman MC, Golub EI, Reddy G, Ward DC, Radding CM, Maizels N. Rad51 expression and localization in B cells carrying out class switch recombination. Proc Natl Acad Sci U S A. 1996 Sep 17;93(19):10222–10227. [PMC free article] [PubMed]
  • Little JB, Nagasawa H. Effect of confluent holding on potentially lethal damage repair, cell cycle progression, and chromosomal aberrations in human normal and ataxia-telangiectasia fibroblasts. Radiat Res. 1985 Jan;101(1):81–93. [PubMed]
  • Löbrich M, Rydberg B, Cooper PK. Repair of x-ray-induced DNA double-strand breaks in specific Not I restriction fragments in human fibroblasts: joining of correct and incorrect ends. Proc Natl Acad Sci U S A. 1995 Dec 19;92(26):12050–12054. [PMC free article] [PubMed]
  • Lukacsovich T, Yang D, Waldman AS. Repair of a specific double-strand break generated within a mammalian chromosome by yeast endonuclease I-SceI. Nucleic Acids Res. 1994 Dec 25;22(25):5649–5657. [PMC free article] [PubMed]
  • Meyn MS. Ataxia-telangiectasia and cellular responses to DNA damage. Cancer Res. 1995 Dec 15;55(24):5991–6001. [PubMed]
  • Meyn MS. High spontaneous intrachromosomal recombination rates in ataxia-telangiectasia. Science. 1993 May 28;260(5112):1327–1330. [PubMed]
  • Miura M, Domon M, Sasaki T, Kondo S, Takasaki Y. Two types of proliferating cell nuclear antigen (PCNA) complex formation in quiescent normal and xeroderma pigmentosum group A fibroblasts following ultraviolet light (uv) irradiation. Exp Cell Res. 1992 Aug;201(2):541–544. [PubMed]
  • Moore JK, Haber JE. Cell cycle and genetic requirements of two pathways of nonhomologous end-joining repair of double-strand breaks in Saccharomyces cerevisiae. Mol Cell Biol. 1996 May;16(5):2164–2173. [PMC free article] [PubMed]
  • Morgan WF, Day JP, Kaplan MI, McGhee EM, Limoli CL. Genomic instability induced by ionizing radiation. Radiat Res. 1996 Sep;146(3):247–258. [PubMed]
  • Murnane JP. Cell cycle regulation in response to DNA damage in mammalian cells: a historical perspective. Cancer Metastasis Rev. 1995 Mar;14(1):17–29. [PubMed]
  • Nevaldine B, Longo JA, King GA, Vilenchik M, Sagerman RH, Hahn PJ. Induction and repair of DNA double-strand breaks. Radiat Res. 1993 Mar;133(3):370–374. [PubMed]
  • Olive PL, Banáth JP, Durand RE. Detection of etoposide resistance by measuring DNA damage in individual Chinese hamster cells. J Natl Cancer Inst. 1990 May 2;82(9):779–783. [PubMed]
  • Olive PL, Wlodek D, Banáth JP. DNA double-strand breaks measured in individual cells subjected to gel electrophoresis. Cancer Res. 1991 Sep 1;51(17):4671–4676. [PubMed]
  • Painter RB. Radioresistant DNA synthesis: an intrinsic feature of ataxia telangiectasia. Mutat Res. 1981 Nov;84(1):183–190. [PubMed]
  • Pandita TK, Hittelman WN. Initial chromosome damage but not DNA damage is greater in ataxia telangiectasia cells. Radiat Res. 1992 Apr;130(1):94–103. [PubMed]
  • Park MS, Knauf JA, Pendergrass SH, Coulon CH, Strniste GF, Marrone BL, MacInnes MA. Ultraviolet-induced movement of the human DNA repair protein, Xeroderma pigmentosum type G, in the nucleus. Proc Natl Acad Sci U S A. 1996 Aug 6;93(16):8368–8373. [PMC free article] [PubMed]
  • Petrini JH, Bressan DA, Yao MS. The RAD52 epistasis group in mammalian double strand break repair. Semin Immunol. 1997 Jun;9(3):181–188. [PubMed]
  • Petrini JH, Walsh ME, DiMare C, Chen XN, Korenberg JR, Weaver DT. Isolation and characterization of the human MRE11 homologue. Genomics. 1995 Sep 1;29(1):80–86. [PubMed]
  • Planchon SM, Wuerzberger S, Frydman B, Witiak DT, Hutson P, Church DR, Wilding G, Boothman DA. Beta-lapachone-mediated apoptosis in human promyelocytic leukemia (HL-60) and human prostate cancer cells: a p53-independent response. Cancer Res. 1995 Sep 1;55(17):3706–3711. [PubMed]
  • Plug AW, Xu J, Reddy G, Golub EI, Ashley T. Presynaptic association of Rad51 protein with selected sites in meiotic chromatin. Proc Natl Acad Sci U S A. 1996 Jun 11;93(12):5920–5924. [PMC free article] [PubMed]
  • Roeder GS. Sex and the single cell: meiosis in yeast. Proc Natl Acad Sci U S A. 1995 Nov 7;92(23):10450–10456. [PMC free article] [PubMed]
  • Saunders WS, Cooke CA, Earnshaw WC. Compartmentalization within the nucleus: discovery of a novel subnuclear region. J Cell Biol. 1991 Nov;115(4):919–931. [PMC free article] [PubMed]
  • Savitsky K, Bar-Shira A, Gilad S, Rotman G, Ziv Y, Vanagaite L, Tagle DA, Smith S, Uziel T, Sfez S, et al. A single ataxia telangiectasia gene with a product similar to PI-3 kinase. Science. 1995 Jun 23;268(5218):1749–1753. [PubMed]
  • Scheffner M, Münger K, Byrne JC, Howley PM. The state of the p53 and retinoblastoma genes in human cervical carcinoma cell lines. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5523–5527. [PMC free article] [PubMed]
  • Schiestl RH, Zhu J, Petes TD. Effect of mutations in genes affecting homologous recombination on restriction enzyme-mediated and illegitimate recombination in Saccharomyces cerevisiae. Mol Cell Biol. 1994 Jul;14(7):4493–4500. [PMC free article] [PubMed]
  • Scully R, Chen J, Plug A, Xiao Y, Weaver D, Feunteun J, Ashley T, Livingston DM. Association of BRCA1 with Rad51 in mitotic and meiotic cells. Cell. 1997 Jan 24;88(2):265–275. [PubMed]
  • Sestili P, Cattabeni F, Cantoni O. Simultaneous determination of DNA double strand breaks and DNA fragment size in cultured mammalian cells exposed to hydrogen peroxide/histidine or etoposide with CHEF electrophoresis. Carcinogenesis. 1995 Apr;16(4):703–706. [PubMed]
  • Shen Z, Denison K, Lobb R, Gatewood JM, Chen DJ. The human and mouse homologs of the yeast RAD52 gene: cDNA cloning, sequence analysis, assignment to human chromosome 12p12.2-p13, and mRNA expression in mouse tissues. Genomics. 1995 Jan 1;25(1):199–206. [PubMed]
  • 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. [PubMed]
  • Shinohara A, Ogawa T. Homologous recombination and the roles of double-strand breaks. Trends Biochem Sci. 1995 Oct;20(10):387–391. [PubMed]
  • Stamato TD, Weinstein R, Giaccia A, Mackenzie L. Isolation of cell cycle-dependent gamma ray-sensitive Chinese hamster ovary cell. Somatic Cell Genet. 1983 Mar;9(2):165–173. [PubMed]
  • Tashiro S, Kotomura N, Shinohara A, Tanaka K, Ueda K, Kamada N. S phase specific formation of the human Rad51 protein nuclear foci in lymphocytes. Oncogene. 1996 May 16;12(10):2165–2170. [PubMed]
  • Toschi L, Bravo R. Changes in cyclin/proliferating cell nuclear antigen distribution during DNA repair synthesis. J Cell Biol. 1988 Nov;107(5):1623–1628. [PMC free article] [PubMed]
  • Tsukamoto Y, Kato J, Ikeda H. Effects of mutations of RAD50, RAD51, RAD52, and related genes on illegitimate recombination in Saccharomyces cerevisiae. Genetics. 1996 Feb;142(2):383–391. [PMC free article] [PubMed]
  • Weaver DT. What to do at an end: DNA double-strand-break repair. Trends Genet. 1995 Oct;11(10):388–392. [PubMed]
  • Weis K, Rambaud S, Lavau C, Jansen J, Carvalho T, Carmo-Fonseca M, Lamond A, Dejean A. Retinoic acid regulates aberrant nuclear localization of PML-RAR alpha in acute promyelocytic leukemia cells. Cell. 1994 Jan 28;76(2):345–356. [PubMed]
  • Yamamoto A, Taki T, Yagi H, Habu T, Yoshida K, Yoshimura Y, Yamamoto K, Matsushiro A, Nishimune Y, Morita T. Cell cycle-dependent expression of the mouse Rad51 gene in proliferating cells. Mol Gen Genet. 1996 Apr 24;251(1):1–12. [PubMed]

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