The effects of AtRad52 over-expression on homologous recombination in Arabidopsis

Aviva Samach, Vyacheslav Gurevich, Naomi Avivi-Ragolsky, Avraham A. Levy

Research output: Contribution to journalArticlepeer-review

Abstract

AtRad52 homologs are involved in DNA recombination and repair, but their precise functions in different homologous recombination (HR) pathways or in gene-targeting have not been analyzed. In order to facilitate our analyses, we generated an AtRad52-1A variant that had a stronger nuclear localization than the native gene thanks to the removal of the transit peptide for mitochondrial localization and to the addition of a nuclear localization signal. Over-expression of this variant increased HR in the nucleus, compared with the native AtRad52-1A: it increased intra-chromosomal recombination and synthesis-dependent strand-annealing HR repair rates; but conversely, it repressed the single-strand annealing pathway. The effect of AtRad52-1A over-expression on gene-targeting was tested with and without the expression of small RNAs generated from an RNAi construct containing homology to the target and donor sequences. True gene-targeting events at the Arabidopsis Cruciferin locus were obtained only when combining AtRad52-1A over-expression and target/donor-specific RNAi. This suggests that sequence-specific small RNAs might be involved in AtRad52-1A-mediated HR.

Original languageEnglish
Pages (from-to)30-40
Number of pages11
JournalPlant Journal
Volume95
Issue number1
Early online date18 Apr 2018
DOIs
StatePublished - Jul 2018

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