Skip to main content
NCBI home page
Genetics logoLink to Genetics
. 2003 Dec;165(4):1831–1842. doi: 10.1093/genetics/165.4.1831

The homologous chromosome is an effective template for the repair of mitotic DNA double-strand breaks in Drosophila.

Yikang S Rong 1, Kent G Golic 1
PMCID: PMC1462885  PMID: 14704169

Abstract

In recombinational DNA double-strand break repair a homologous template for gene conversion may be located at several different genomic positions: on the homologous chromosome in diploid organisms, on the sister chromatid after DNA replication, or at an ectopic position. The use of the homologous chromosome in mitotic gene conversion is thought to be limited in the yeast Saccharomyces cerevisiae and mammalian cells. In contrast, by studying the repair of double-strand breaks generated by the I-SceI rare-cutting endonuclease, we find that the homologous chromosome is frequently used in Drosophila melanogaster, which we suggest is attributable to somatic pairing of homologous chromosomes in mitotic cells of Drosophila. We also find that Drosophila mitotic cells of the germ line, like yeast, employ the homologous recombinational repair pathway more often than imperfect nonhomologous end joining.

Full Text

The Full Text of this article is available as a PDF (223.3 KB).

Selected References

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

  1. Adams Melissa D., McVey Mitch, Sekelsky Jeff J. Drosophila BLM in double-strand break repair by synthesis-dependent strand annealing. Science. 2003 Jan 10;299(5604):265–267. doi: 10.1126/science.1077198. [DOI] [PubMed] [Google Scholar]
  2. Beall E. L., Rio D. C. Drosophila P-element transposase is a novel site-specific endonuclease. Genes Dev. 1997 Aug 15;11(16):2137–2151. doi: 10.1101/gad.11.16.2137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bellaiche Y., Mogila V., Perrimon N. I-SceI endonuclease, a new tool for studying DNA double-strand break repair mechanisms in Drosophila. Genetics. 1999 Jul;152(3):1037–1044. doi: 10.1093/genetics/152.3.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bibikova Marina, Golic Mary, Golic Kent G., Carroll Dana. Targeted chromosomal cleavage and mutagenesis in Drosophila using zinc-finger nucleases. Genetics. 2002 Jul;161(3):1169–1175. doi: 10.1093/genetics/161.3.1169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burgess S. M., Kleckner N., Weiner B. M. Somatic pairing of homologs in budding yeast: existence and modulation. Genes Dev. 1999 Jun 15;13(12):1627–1641. doi: 10.1101/gad.13.12.1627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Colleaux L., D'Auriol L., Galibert F., Dujon B. Recognition and cleavage site of the intron-encoded omega transposase. Proc Natl Acad Sci U S A. 1988 Aug;85(16):6022–6026. doi: 10.1073/pnas.85.16.6022. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Donoho G., Jasin M., Berg P. Analysis of gene targeting and intrachromosomal homologous recombination stimulated by genomic double-strand breaks in mouse embryonic stem cells. Mol Cell Biol. 1998 Jul;18(7):4070–4078. doi: 10.1128/mcb.18.7.4070. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ducau J., Bregliano J. C., de La Roche Saint-André C. Gamma-irradiation stimulates homology-directed DNA double-strand break repair in Drosophila embryo. Mutat Res. 2000 Jun 30;460(1):69–80. doi: 10.1016/s0921-8777(00)00017-3. [DOI] [PubMed] [Google Scholar]
  9. Engels W. R., Johnson-Schlitz D. M., Eggleston W. B., Sved J. High-frequency P element loss in Drosophila is homolog dependent. Cell. 1990 Aug 10;62(3):515–525. doi: 10.1016/0092-8674(90)90016-8. [DOI] [PubMed] [Google Scholar]
  10. Engels W. R., Preston C. R., Johnson-Schlitz D. M. Long-range cis preference in DNA homology search over the length of a Drosophila chromosome. Science. 1994 Mar 18;263(5153):1623–1625. doi: 10.1126/science.8128250. [DOI] [PubMed] [Google Scholar]
  11. Esposito M. S. Evidence that spontaneous mitotic recombination occurs at the two-strand stage. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4436–4440. doi: 10.1073/pnas.75.9.4436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fabre F. Induced intragenic recombination in yeast can occur during the G1 mitotic phase. Nature. 1978 Apr 27;272(5656):795–798. doi: 10.1038/272795a0. [DOI] [PubMed] [Google Scholar]
  13. Fung J. C., Marshall W. F., Dernburg A., Agard D. A., Sedat J. W. Homologous chromosome pairing in Drosophila melanogaster proceeds through multiple independent initiations. J Cell Biol. 1998 Apr 6;141(1):5–20. doi: 10.1083/jcb.141.1.5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gloor G. B., Preston C. R., Johnson-Schlitz D. M., Nassif N. A., Phillis R. W., Benz W. K., Robertson H. M., Engels W. R. Type I repressors of P element mobility. Genetics. 1993 Sep;135(1):81–95. doi: 10.1093/genetics/135.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Golic M. M., Golic K. G. A quantitative measure of the mitotic pairing of alleles in Drosophila melanogaster and the influence of structural heterozygosity. Genetics. 1996 May;143(1):385–400. doi: 10.1093/genetics/143.1.385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gong Wei J., Golic Kent G. Ends-out, or replacement, gene targeting in Drosophila. Proc Natl Acad Sci U S A. 2003 Feb 14;100(5):2556–2561. doi: 10.1073/pnas.0535280100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hiraoka Y., Dernburg A. F., Parmelee S. J., Rykowski M. C., Agard D. A., Sedat J. W. The onset of homologous chromosome pairing during Drosophila melanogaster embryogenesis. J Cell Biol. 1993 Feb;120(3):591–600. doi: 10.1083/jcb.120.3.591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jeggo P. A. DNA breakage and repair. Adv Genet. 1998;38:185–218. doi: 10.1016/s0065-2660(08)60144-3. [DOI] [PubMed] [Google Scholar]
  19. Johnson R. D., Jasin M. Sister chromatid gene conversion is a prominent double-strand break repair pathway in mammalian cells. EMBO J. 2000 Jul 3;19(13):3398–3407. doi: 10.1093/emboj/19.13.3398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kadyk L. C., Hartwell L. H. Sister chromatids are preferred over homologs as substrates for recombinational repair in Saccharomyces cerevisiae. Genetics. 1992 Oct;132(2):387–402. doi: 10.1093/genetics/132.2.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lee S. E., Pâques F., Sylvan J., Haber J. E. Role of yeast SIR genes and mating type in directing DNA double-strand breaks to homologous and non-homologous repair paths. Curr Biol. 1999 Jul 15;9(14):767–770. doi: 10.1016/s0960-9822(99)80339-x. [DOI] [PubMed] [Google Scholar]
  22. Liang F., Han M., Romanienko P. J., Jasin M. Homology-directed repair is a major double-strand break repair pathway in mammalian cells. Proc Natl Acad Sci U S A. 1998 Apr 28;95(9):5172–5177. doi: 10.1073/pnas.95.9.5172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lin F. L., Sperle K., Sternberg N. Model for homologous recombination during transfer of DNA into mouse L cells: role for DNA ends in the recombination process. Mol Cell Biol. 1984 Jun;4(6):1020–1034. doi: 10.1128/mcb.4.6.1020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lin Y., Lukacsovich T., Waldman A. S. Multiple pathways for repair of DNA double-strand breaks in mammalian chromosomes. Mol Cell Biol. 1999 Dec;19(12):8353–8360. doi: 10.1128/mcb.19.12.8353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Malkova A., Ivanov E. L., Haber J. E. Double-strand break repair in the absence of RAD51 in yeast: a possible role for break-induced DNA replication. Proc Natl Acad Sci U S A. 1996 Jul 9;93(14):7131–7136. doi: 10.1073/pnas.93.14.7131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Maryon E., Carroll D. Characterization of recombination intermediates from DNA injected into Xenopus laevis oocytes: evidence for a nonconservative mechanism of homologous recombination. Mol Cell Biol. 1991 Jun;11(6):3278–3287. doi: 10.1128/mcb.11.6.3278. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Moynahan M. E., Jasin M. Loss of heterozygosity induced by a chromosomal double-strand break. Proc Natl Acad Sci U S A. 1997 Aug 19;94(17):8988–8993. doi: 10.1073/pnas.94.17.8988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Perrin A., Buckle M., Dujon B. Asymmetrical recognition and activity of the I-SceI endonuclease on its site and on intron-exon junctions. EMBO J. 1993 Jul;12(7):2939–2947. doi: 10.1002/j.1460-2075.1993.tb05956.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Plessis A., Perrin A., Haber J. E., Dujon B. Site-specific recombination determined by I-SceI, a mitochondrial group I intron-encoded endonuclease expressed in the yeast nucleus. Genetics. 1992 Mar;130(3):451–460. doi: 10.1093/genetics/130.3.451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Preston Christine R., Engels William, Flores Carlos. Efficient repair of DNA breaks in Drosophila: evidence for single-strand annealing and competition with other repair pathways. Genetics. 2002 Jun;161(2):711–720. doi: 10.1093/genetics/161.2.711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Puchta H., Dujon B., Hohn B. Homologous recombination in plant cells is enhanced by in vivo induction of double strand breaks into DNA by a site-specific endonuclease. Nucleic Acids Res. 1993 Nov 11;21(22):5034–5040. doi: 10.1093/nar/21.22.5034. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Pâques F., Haber J. E. Multiple pathways of recombination induced by double-strand breaks in Saccharomyces cerevisiae. Microbiol Mol Biol Rev. 1999 Jun;63(2):349–404. doi: 10.1128/mmbr.63.2.349-404.1999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Richardson C., Moynahan M. E., Jasin M. Double-strand break repair by interchromosomal recombination: suppression of chromosomal translocations. Genes Dev. 1998 Dec 15;12(24):3831–3842. doi: 10.1101/gad.12.24.3831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rong Y. S., Golic K. G. Dominant defects in Drosophila eye pigmentation resulting from a euchromatin-heterochromatin fusion gene. Genetics. 1998 Dec;150(4):1551–1566. doi: 10.1093/genetics/150.4.1551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Rong Y. S., Golic K. G. Gene targeting by homologous recombination in Drosophila. Science. 2000 Jun 16;288(5473):2013–2018. doi: 10.1126/science.288.5473.2013. [DOI] [PubMed] [Google Scholar]
  36. Rong Yikang S., Titen Simon W., Xie Heng B., Golic Mary M., Bastiani Michael, Bandyopadhyay Pradip, Olivera Baldomero M., Brodsky Michael, Rubin Gerald M., Golic Kent G. Targeted mutagenesis by homologous recombination in D. melanogaster. Genes Dev. 2002 Jun 15;16(12):1568–1581. doi: 10.1101/gad.986602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Rouet P., Smih F., Jasin M. Expression of a site-specific endonuclease stimulates homologous recombination in mammalian cells. Proc Natl Acad Sci U S A. 1994 Jun 21;91(13):6064–6068. doi: 10.1073/pnas.91.13.6064. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sjögren C., Nasmyth K. Sister chromatid cohesion is required for postreplicative double-strand break repair in Saccharomyces cerevisiae. Curr Biol. 2001 Jun 26;11(12):991–995. doi: 10.1016/s0960-9822(01)00271-8. [DOI] [PubMed] [Google Scholar]
  39. Stark Jeremy M., Jasin Maria. Extensive loss of heterozygosity is suppressed during homologous repair of chromosomal breaks. Mol Cell Biol. 2003 Jan;23(2):733–743. doi: 10.1128/MCB.23.2.733-743.2003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Takata M., Sasaki M. S., Sonoda E., Morrison C., Hashimoto M., Utsumi H., Yamaguchi-Iwai Y., Shinohara A., Takeda S. Homologous recombination and non-homologous end-joining pathways of DNA double-strand break repair have overlapping roles in the maintenance of chromosomal integrity in vertebrate cells. EMBO J. 1998 Sep 15;17(18):5497–5508. doi: 10.1093/emboj/17.18.5497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Virgin J. B., Bailey J. P., Hasteh F., Neville J., Cole A., Tromp G. Crossing over is rarely associated with mitotic intragenic recombination in Schizosaccharomyces pombe. Genetics. 2001 Jan;157(1):63–77. doi: 10.1093/genetics/157.1.63. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES