Skip to main content
NCBI home page
The Plant Cell logoLink to The Plant Cell
. 1996 Nov;8(11):2057–2066. doi: 10.1105/tpc.8.11.2057

Enhancement of somatic intrachromosomal homologous recombination in Arabidopsis by the HO endonuclease.

M Chiurazzi 1, A Ray 1, J F Viret 1, R Perera 1, X H Wang 1, A M Lloyd 1, E R Signer 1
PMCID: PMC161334  PMID: 8953770

Abstract

The HO endonuclease promotes gene conversion between mating-type alleles in yeast by a DNA double-strand break at the site of conversion (the MAT-Y/Z site). As a first step toward understanding the molecular basis of homologous recombination in higher plants, we demonstrate that expression of HO in Arabidopsis enhances intrachromosomal recombination between inverted repeats of two defective beta-glucuronidase (gus) genes (GUS- test construct). One of these genes has the Y/Z site. The two genes share 2.5 kb of DNA sequence homology around the HO cut site. Somatic recombination between the two repeats was determined by using a histochemical assay of GUS activity. The frequency of Gus+ sectors in leaves of F1 plants from a cross between parents homozygous for the GUS- test construct and HO, respectively, was 10-fold higher than in F1 plants from a cross between the same plant containing the GUS- test construct and a wild-type parent. Polymerase chain reaction analysis showed restoration of the 5' end of the GUS gene in recombinant sectors. The induction of intrachromosomal gene conversion in Arabidopsis by HO reveals the general utility of site-specific DNA endonucleases in producing targeted homologous recombination in plant genomes.

Full Text

The Full Text of this article is available as a PDF (1.8 MB).

Selected References

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

  1. Assaad F. F., Signer E. R. Somatic and germinal recombination of a direct repeat in Arabidopsis. Genetics. 1992 Oct;132(2):553–566. doi: 10.1093/genetics/132.2.553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Athma P., Peterson T. Ac induces homologous recombination at the maize P locus. Genetics. 1991 May;128(1):163–173. doi: 10.1093/genetics/128.1.163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cao L., Alani E., Kleckner N. A pathway for generation and processing of double-strand breaks during meiotic recombination in S. cerevisiae. Cell. 1990 Jun 15;61(6):1089–1101. doi: 10.1016/0092-8674(90)90072-m. [DOI] [PubMed] [Google Scholar]
  4. Das O. P., Levi-Minzi S., Koury M., Benner M., Messing J. A somatic gene rearrangement contributing to genetic diversity in maize. Proc Natl Acad Sci U S A. 1990 Oct;87(20):7809–7813. doi: 10.1073/pnas.87.20.7809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Davies C., Howard D., Tam G., Wong N. Isolation of Arabidopsis thaliana mutants hypersensitive to gamma radiation. Mol Gen Genet. 1994 Jun 15;243(6):660–665. doi: 10.1007/BF00279575. [DOI] [PubMed] [Google Scholar]
  6. Flavell R. B. Inactivation of gene expression in plants as a consequence of specific sequence duplication. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3490–3496. doi: 10.1073/pnas.91.9.3490. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gloor G. B., Nassif N. A., Johnson-Schlitz D. M., Preston C. R., Engels W. R. Targeted gene replacement in Drosophila via P element-induced gap repair. Science. 1991 Sep 6;253(5024):1110–1117. doi: 10.1126/science.1653452. [DOI] [PubMed] [Google Scholar]
  8. Haber J. E. In vivo biochemistry: physical monitoring of recombination induced by site-specific endonucleases. Bioessays. 1995 Jul;17(7):609–620. doi: 10.1002/bies.950170707. [DOI] [PubMed] [Google Scholar]
  9. Haber J. E., Ray B. L., Kolb J. M., White C. I. Rapid kinetics of mismatch repair of heteroduplex DNA that is formed during recombination in yeast. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3363–3367. doi: 10.1073/pnas.90.8.3363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hastings P. J., McGill C., Shafer B., Strathern J. N. Ends-in vs. ends-out recombination in yeast. Genetics. 1993 Dec;135(4):973–980. doi: 10.1093/genetics/135.4.973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Herskowitz I., Jensen R. E. Putting the HO gene to work: practical uses for mating-type switching. Methods Enzymol. 1991;194:132–146. doi: 10.1016/0076-6879(91)94011-z. [DOI] [PubMed] [Google Scholar]
  12. Jackson J. A., Fink G. R. Gene conversion between duplicated genetic elements in yeast. Nature. 1981 Jul 23;292(5821):306–311. doi: 10.1038/292306a0. [DOI] [PubMed] [Google Scholar]
  13. Jefferson R. A., Kavanagh T. A., Bevan M. W. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 1987 Dec 20;6(13):3901–3907. doi: 10.1002/j.1460-2075.1987.tb02730.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Klein H. L. Lack of association between intrachromosomal gene conversion and reciprocal exchange. 1984 Aug 30-Sep 5Nature. 310(5980):748–753. doi: 10.1038/310748a0. [DOI] [PubMed] [Google Scholar]
  15. Kobayashi I. Mechanisms for gene conversion and homologous recombination: the double-strand break repair model and the successive half crossing-over model. Adv Biophys. 1992;28:81–133. doi: 10.1016/0065-227x(92)90023-k. [DOI] [PubMed] [Google Scholar]
  16. Kolodkin A. L., Klar A. J., Stahl F. W. Double-strand breaks can initiate meiotic recombination in S. cerevisiae. Cell. 1986 Aug 29;46(5):733–740. doi: 10.1016/0092-8674(86)90349-1. [DOI] [PubMed] [Google Scholar]
  17. Kostriken R., Strathern J. N., Klar A. J., Hicks J. B., Heffron F. A site-specific endonuclease essential for mating-type switching in Saccharomyces cerevisiae. Cell. 1983 Nov;35(1):167–174. doi: 10.1016/0092-8674(83)90219-2. [DOI] [PubMed] [Google Scholar]
  18. Lazo G. R., Stein P. A., Ludwig R. A. A DNA transformation-competent Arabidopsis genomic library in Agrobacterium. Biotechnology (N Y) 1991 Oct;9(10):963–967. doi: 10.1038/nbt1091-963. [DOI] [PubMed] [Google Scholar]
  19. Lin F. L., Sperle K., Sternberg N. Intermolecular recombination between DNAs introduced into mouse L cells is mediated by a nonconservative pathway that leads to crossover products. Mol Cell Biol. 1990 Jan;10(1):103–112. doi: 10.1128/mcb.10.1.103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lloyd A. M., Walbot V., Davis R. W. Arabidopsis and Nicotiana anthocyanin production activated by maize regulators R and C1. Science. 1992 Dec 11;258(5089):1773–1775. doi: 10.1126/science.1465611. [DOI] [PubMed] [Google Scholar]
  21. Marsh J. L., Erfle M., Wykes E. J. The pIC plasmid and phage vectors with versatile cloning sites for recombinant selection by insertional inactivation. Gene. 1984 Dec;32(3):481–485. doi: 10.1016/0378-1119(84)90022-2. [DOI] [PubMed] [Google Scholar]
  22. Mittelsten Scheid O., Paszkowski J., Potrykus I. Reversible inactivation of a transgene in Arabidopsis thaliana. Mol Gen Genet. 1991 Aug;228(1-2):104–112. doi: 10.1007/BF00282454. [DOI] [PubMed] [Google Scholar]
  23. Myers R. S., Stahl F. W. Chi and the RecBC D enzyme of Escherichia coli. Annu Rev Genet. 1994;28:49–70. doi: 10.1146/annurev.ge.28.120194.000405. [DOI] [PubMed] [Google Scholar]
  24. Nassif N., Penney J., Pal S., Engels W. R., Gloor G. B. Efficient copying of nonhomologous sequences from ectopic sites via P-element-induced gap repair. Mol Cell Biol. 1994 Mar;14(3):1613–1625. doi: 10.1128/mcb.14.3.1613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nickoloff J. A., Chen E. Y., Heffron F. A 24-base-pair DNA sequence from the MAT locus stimulates intergenic recombination in yeast. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7831–7835. doi: 10.1073/pnas.83.20.7831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pelham H. R. A regulatory upstream promoter element in the Drosophila hsp 70 heat-shock gene. Cell. 1982 Sep;30(2):517–528. doi: 10.1016/0092-8674(82)90249-5. [DOI] [PubMed] [Google Scholar]
  27. Perez P., Tiraby G., Kallerhoff J., Perret J. Phleomycin resistance as a dominant selectable marker for plant cell transformation. Plant Mol Biol. 1989 Oct;13(4):365–373. doi: 10.1007/BF00015548. [DOI] [PubMed] [Google Scholar]
  28. Petes T. D., Hill C. W. Recombination between repeated genes in microorganisms. Annu Rev Genet. 1988;22:147–168. doi: 10.1146/annurev.ge.22.120188.001051. [DOI] [PubMed] [Google Scholar]
  29. Puchta H., Swoboda P., Gal S., Blot M., Hohn B. Somatic intrachromosomal homologous recombination events in populations of plant siblings. Plant Mol Biol. 1995 May;28(2):281–292. doi: 10.1007/BF00020247. [DOI] [PubMed] [Google Scholar]
  30. Ray A., Machin N., Stahl F. W. A DNA double chain break stimulates triparental recombination in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6225–6229. doi: 10.1073/pnas.86.16.6225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ray A., Siddiqi I., Kolodkin A. L., Stahl F. W. Intra-chromosomal gene conversion induced by a DNA double-strand break in Saccharomyces cerevisiae. J Mol Biol. 1988 May 20;201(2):247–260. doi: 10.1016/0022-2836(88)90136-2. [DOI] [PubMed] [Google Scholar]
  32. Rouet P., Smih F., Jasin M. Introduction of double-strand breaks into the genome of mouse cells by expression of a rare-cutting endonuclease. Mol Cell Biol. 1994 Dec;14(12):8096–8106. doi: 10.1128/mcb.14.12.8096. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rudin N., Sugarman E., Haber J. E. Genetic and physical analysis of double-strand break repair and recombination in Saccharomyces cerevisiae. Genetics. 1989 Jul;122(3):519–534. doi: 10.1093/genetics/122.3.519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Russell D. W., Jensen R., Zoller M. J., Burke J., Errede B., Smith M., Herskowitz I. Structure of the Saccharomyces cerevisiae HO gene and analysis of its upstream regulatory region. Mol Cell Biol. 1986 Dec;6(12):4281–4294. doi: 10.1128/mcb.6.12.4281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Segall A. M., Roth J. R. Approaches to half-tetrad analysis in bacteria: recombination between repeated, inverse-order chromosomal sequences. Genetics. 1994 Jan;136(1):27–39. doi: 10.1093/genetics/136.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Steller H., Pirrotta V. Regulated expression of genes injected into early Drosophila embryos. EMBO J. 1984 Jan;3(1):165–173. doi: 10.1002/j.1460-2075.1984.tb01778.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Strathern J. N., Klar A. J., Hicks J. B., Abraham J. A., Ivy J. M., Nasmyth K. A., McGill C. Homothallic switching of yeast mating type cassettes is initiated by a double-stranded cut in the MAT locus. Cell. 1982 Nov;31(1):183–192. doi: 10.1016/0092-8674(82)90418-4. [DOI] [PubMed] [Google Scholar]
  38. Sun H., Treco D., Schultes N. P., Szostak J. W. Double-strand breaks at an initiation site for meiotic gene conversion. Nature. 1989 Mar 2;338(6210):87–90. doi: 10.1038/338087a0. [DOI] [PubMed] [Google Scholar]
  39. Szostak J. W., Orr-Weaver T. L., Rothstein R. J., Stahl F. W. The double-strand-break repair model for recombination. Cell. 1983 May;33(1):25–35. doi: 10.1016/0092-8674(83)90331-8. [DOI] [PubMed] [Google Scholar]
  40. Tovar J., Lichtenstein C. Somatic and Meiotic Chromosomal Recombination between Inverted Duplications in Transgenic Tobacco Plants. Plant Cell. 1992 Mar;4(3):319–332. doi: 10.1105/tpc.4.3.319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wing D., Koncz C., Schell J. Conserved function in Nicotiana tabacum of a single Drosophila hsp70 promoter heat shock element when fused to a minimal T-DNA promoter. Mol Gen Genet. 1989 Oct;219(1-2):9–16. doi: 10.1007/BF00261151. [DOI] [PubMed] [Google Scholar]
  42. Wu T. C., Lichten M. Meiosis-induced double-strand break sites determined by yeast chromatin structure. Science. 1994 Jan 28;263(5146):515–518. doi: 10.1126/science.8290959. [DOI] [PubMed] [Google Scholar]
  43. de Massy B., Rocco V., Nicolas A. The nucleotide mapping of DNA double-strand breaks at the CYS3 initiation site of meiotic recombination in Saccharomyces cerevisiae. EMBO J. 1995 Sep 15;14(18):4589–4598. doi: 10.1002/j.1460-2075.1995.tb00138.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Plant Cell are provided here courtesy of Oxford University Press

RESOURCES