Journal Article

Non-homologous DNA end joining in plant cells is associated with deletions and filler DNA insertions

Vera Gorbunova and Avraham A. Levy

in Nucleic Acids Research

Volume 25, issue 22, pages 4650-4657
Published in print November 1997 | ISSN: 0305-1048
Published online November 1997 | e-ISSN: 1362-4962 | DOI: https://dx.doi.org/10.1093/nar/25.22.4650
Non-homologous DNA end joining in plant cells is associated with deletions and filler DNA insertions

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Double strand DNA breaks in plants are primarily repaired via non-homologous end joining. However, little is known about the molecular events underlying this process. We have studied non-homologous end joining of linearized plasmid DNA with different termini configurations following transformation into tobacco cells. A variety of sequences were found at novel end junctions. Joining with no sequence alterations was rare. In most cases, deletions were found at both ends, and rejoining usually occurred at short repeats. A distinct feature of plant junctions was the presence of relatively large, up to 1.2 kb long, insertions (filler DNA), in ∼30% of the analyzed clones. The filler DNA originated either from internal regions of the plasmid or from tobacco genomic DNA. Some insertions had a complex structure consisting of several reshuffled plasmid-related regions. These data suggest that double strand break repair in plants involves extensive end degradation, DNA synthesis following invasion of ectopic templates and multiple template switches. Such a mechanism is reminiscent of the synthesis-dependent recombination in bacteriophage T4. It can also explain the frequent ‘DNA scrambling’ associated with illegitimate recombination in plants.

Journal Article.  4419 words.  Illustrated.

Subjects: Chemistry ; Biochemistry ; Bioinformatics and Computational Biology ; Genetics and Genomics ; Molecular and Cell Biology

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