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PMID: 8643528 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Two different but related mechanisms are used in plants for the repair of genomic double-strand breaks by homologous recombination.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 93 ·No. 10 ·1996-05-14 ·Pages 5055-60

Puchta H, Dujon B, Hohn B

Abstract

Genomic double-strand breaks (DSBs) are key intermediates in recombination reactions of living organisms. We studied the repair of genomic DSBs by homologous sequences in plants. Tobacco plants containing a site for the highly specific restriction enzyme I-Sce I were cotransformed with Agrobacterium strains carrying sequences homologous to the transgene locus and, separately, containing the gene coding for the enzyme. We show that the induction of a DSB can increase the frequency of homologous recombination at a specific locus by up to two orders of magnitude. Analysis of the recombination products demonstrates that a DSB can be repaired via homologous recombination by at least two different but related pathways. In the major pathway, homologies on both sides of the DSB are used, analogous to the conservative DSB repair model originally proposed for meiotic recombination in yeast. Homologous recombination of the minor pathway is restricted to one side of the DSB as described by the nonconservative one-sided invasion model. The sequence of the recombination partners was absolutely conserved in two cases, whereas in a third case, a deletion of 14 bp had occurred, probably due to DNA polymerase slippage during the copy process. The induction of DSB breaks to enhance homologous recombination can be applied for a variety of approaches of plant genome manipulation.

MeSH Terms
Base Sequence Cloning, Molecular DNA Damage DNA Repair/genetics DNA, Plant/genetics,metabolism Deoxyribonucleases, Type II Site-Specific/metabolism Gene Targeting Genome, Plant Models, Genetic Molecular Sequence Data Plants/genetics,metabolism,microbiology Plants, Genetically Modified Plants, Toxic Polymerase Chain Reaction Recombination, Genetic Rhizobium/genetics Saccharomyces cerevisiae Proteins Sequence Deletion Tobacco/genetics,metabolism,microbiology Transformation, Genetic
Chemicals
DNA, Plant Saccharomyces cerevisiae Proteins SCEI protein, S cerevisiae Deoxyribonucleases, Type II Site-Specific
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Puchta H
Friedrich Miescher-Institut, Basel, Switzerland.
Dujon B
Hohn B
References (48)
48 references, click to expand
  1. 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):5649-57 PMID: 7838718
  2. Extrachromosomal homologous DNA recombination in plant cells is fast and is not affected by CpG methylation.
    Mol Cell Biol. 1992 Aug;12(8):3372-9 PMID: 1630452
  3. Nested chromosomal fragmentation in yeast using the meganuclease I-Sce I: a new method for physical mapping of eukaryotic genomes.
    Nucleic Acids Res. 1992 Nov 11;20(21):5625-31 PMID: 1333585
  4. Site-specific recombination determined by I-SceI, a mitochondrial group I intron-encoded endonuclease expressed in the yeast nucleus.
    Genetics. 1992 Mar;130(3):451-60 PMID: 1551570
  5. DNA synthesis errors associated with double-strand-break repair.
    Genetics. 1995 Jul;140(3):965-72 PMID: 7672595
  6. 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):1020-34 PMID: 6330525
  7. Asymmetrical recognition and activity of the I-SceI endonuclease on its site and on intron-exon junctions.
    EMBO J. 1993 Jul;12(7):2939-47 PMID: 8335007
  8. Nonreciprocal homologous recombination between Agrobacterium transferred DNA and a plant chromosomal locus.
    Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):7346-50 PMID: 8346254
  9. Cre recombinase-mediated site-specific recombination between plant chromosomes.
    Proc Natl Acad Sci U S A. 1994 Mar 1;91(5):1706-10 PMID: 8127869
  10. Meiosis-induced double-strand break sites determined by yeast chromatin structure.
    Science. 1994 Jan 28;263(5146):515-8 PMID: 8290959
  11. Expression in transgenic tobacco of the bacterial neomycin phosphotransferase gene modified by intron insertions of various sizes.
    Plant Mol Biol. 1992 Aug;19(5):825-36 PMID: 1322741
  12. One-sided invasion events in homologous recombination at double-strand breaks.
    Mutat Res. 1994 May;314(3):199-208 PMID: 7513053
  13. Stress-induced intrachromosomal recombination in plant somatic cells.
    Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):422-6 PMID: 11607349
  14. Homology-associated nonhomologous recombination in mammalian gene targeting.
    Proc Natl Acad Sci U S A. 1994 Aug 30;91(18):8527-31 PMID: 8078916
  15. Mating-type gene switching in Saccharomyces cerevisiae.
    Trends Genet. 1992 Dec;8(12):446-52 PMID: 1492369
  16. Altering the genome by homologous recombination.
    Science. 1989 Jun 16;244(4910):1288-92 PMID: 2660260
  17. Ac induces homologous recombination at the maize P locus.
    Genetics. 1991 May;128(1):163-73 PMID: 1648001
  18. Recombination between prokaryotic and eukaryotic DNA: integration of Agrobacterium tumefaciens T-DNA into the plant genome.
    Genet Eng (N Y). 1995;17:209-29 PMID: 7779513
  19. Somatic and Meiotic Chromosomal Recombination between Inverted Duplications in Transgenic Tobacco Plants.
    Plant Cell. 1992 Mar;4(3):319-332 PMID: 12297648
  20. Targeted recombination with single-stranded DNA vectors in mammalian cells.
    Nucleic Acids Res. 1993 Feb 11;21(3):407-12 PMID: 8441653
  21. Gene targeting in Arabidopsis thaliana.
    Mol Gen Genet. 1992 Jan;231(2):186-93 PMID: 1310519
  22. Active Mutator elements suppress the knotted phenotype and increase recombination at the Kn1-O tandem duplication.
    Genetics. 1992 Nov;132(3):813-22 PMID: 1334895
  23. Induction of homologous recombination in mammalian chromosomes by using the I-SceI system of Saccharomyces cerevisiae.
    Mol Cell Biol. 1995 Apr;15(4):1968-73 PMID: 7891691
  24. Site-specific integration of DNA into wild-type and mutant lox sites placed in the plant genome.
    Plant J. 1995 Apr;7(4):649-59 PMID: 7742860
  25. High fidelity extrachromosomal recombination and gene targeting in plants.
    Mol Gen Genet. 1994 Apr;243(1):106-11 PMID: 8190063
  26. Binary Agrobacterium vectors for plant transformation.
    Nucleic Acids Res. 1984 Nov 26;12(22):8711-21 PMID: 6095209
  27. A system for insertional mutagenesis and chromosomal rearrangement using the Ds transposon and Cre-lox.
    Plant J. 1995 Apr;7(4):687-701 PMID: 7742862
  28. 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):5034-40 PMID: 8255757
  29. In vivo biochemistry: physical monitoring of recombination induced by site-specific endonucleases.
    Bioessays. 1995 Jul;17(7):609-20 PMID: 7646483
  30. The complete sequence of the 8.2 kb segment left of MAT on chromosome III reveals five ORFs, including a gene for a yeast ribokinase.
    Yeast. 1990 Nov-Dec;6(6):521-34 PMID: 1964349
  31. Agrobacterium tumefaciens transfers single-stranded transferred DNA (T-DNA) into the plant cell nucleus.
    Proc Natl Acad Sci U S A. 1994 Aug 16;91(17):8000-4 PMID: 11607492
  32. Alteration of gene expression by restriction enzymes electroporated into plant cells.
    Mutat Res. 1993 Jun;302(2):75-82 PMID: 7684508
  33. Gene targeting in plants.
    EMBO J. 1988 Dec 20;7(13):4021-6 PMID: 16453864
  34. The Agrobacterium tumefaciens virulence D2 protein is responsible for precise integration of T-DNA into the plant genome.
    EMBO J. 1995 Jul 17;14(14):3585-95 PMID: 7628458
  35. Targeted recombination in plants using Agrobacterium coincides with additional rearrangements at the target locus.
    Plant J. 1995 Jan;7(1):109-19 PMID: 7894502
  36. Intrachromosomal homologous recombination in whole plants.
    EMBO J. 1994 Jan 15;13(2):484-9 PMID: 8313893
  37. The double-strand-break repair model for recombination.
    Cell. 1983 May;33(1):25-35 PMID: 6380756
  38. Extrachromosomal homologous recombination and gene targeting in plant cells after Agrobacterium mediated transformation.
    EMBO J. 1990 Oct;9(10):3077-84 PMID: 2209538
  39. Homologous recombination in plant cells after Agrobacterium-mediated transformation.
    Plant Cell. 1990 May;2(5):415-25 PMID: 2152167
  40. Improved binary vectors for Agrobacterium-mediated plant transformation.
    Plant Mol Biol. 1990 Feb;14(2):269-76 PMID: 1966276
  41. Recognition and cleavage site of the intron-encoded omega transposase.
    Proc Natl Acad Sci U S A. 1988 Aug;85(16):6022-6 PMID: 2842757
  42. Targeted disruption of the TGA3 locus in Arabidopsis thaliana.
    Plant J. 1995 Feb;7(2):359-65 PMID: 7704051
  43. Expression of a site-specific endonuclease stimulates homologous recombination in mammalian cells.
    Proc Natl Acad Sci U S A. 1994 Jun 21;91(13):6064-8 PMID: 8016116
  44. Endonuclease-induced, targeted homologous extrachromosomal recombination in Xenopus oocytes.
    Proc Natl Acad Sci U S A. 1995 Jan 31;92(3):806-10 PMID: 7846056
  45. Interference of DNA sequence divergence with precise recombinational DNA repair in mammalian cells.
    EMBO J. 1994 Nov 15;13(22):5355-60 PMID: 7957101
  46. Intra-chromosomal rearrangements generated by Cre-lox site-specific recombination.
    Nucleic Acids Res. 1995 Feb 11;23(3):485-90 PMID: 7885845
  47. 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):8096-106 PMID: 7969147
  48. Gene replacement with one-sided homologous recombination.
    Mol Cell Biol. 1992 Jan;12(1):360-7 PMID: 1729610
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.93.10.5055
Published
1996-05-14
Pages
5055-60
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC39405
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.93.10.5055 PMC Full Text View journal details
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