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Poly(ADP-ribose)polymerases are involved in microhomology mediated back-up non-homologous end joining in Arabidopsis thaliana

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Abstract

Besides the KU-dependent classical non-homologous end-joining (C-NHEJ) pathway, an alternative NHEJ pathway first identified in mammalian systems, which is often called the back-up NHEJ (B-NHEJ) pathway, was also found in plants. In mammalian systems PARP was found to be one of the essential components in B-NHEJ. Here we investigated whether PARP1 and PARP2 were also involved in B-NHEJ in Arabidopsis. To this end Arabidopsis parp1, parp2 and parp1parp2 (p1p2) mutants were isolated and functionally characterized. The p1p2 double mutant was crossed with the C-NHEJ ku80 mutant resulting in the parp1parp2ku80 (p1p2k80) triple mutant. As expected, because of their role in single strand break repair (SSBR) and base excision repair (BER), the p1p2 and p1p2k80 mutants were shown to be sensitive to treatment with the DNA damaging agent MMS. End-joining assays in cell-free leaf protein extracts of the different mutants using linear DNA substrates with different ends reflecting a variety of double strand breaks were performed. The results showed that compatible 5′-overhangs were accurately joined in all mutants, that KU80 protected the ends preventing the formation of large deletions and that PARP proteins were involved in microhomology mediated end joining (MMEJ), one of the characteristics of B-NHEJ.

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Acknowledgments

We thank Dr. Li Liang for providing the plasmid PUC19PD1/4. This work was financially supported by the Chinese Scholarship Council (CSC) (QJ, HS) and the European Union Program EU Recbreed (KBBE-2008-227190) (SdP).

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  1. Qi Jia

    Present address: College of Crop Science, Fujian Agriculture and Forestry University, Fuzhou, China

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  1. Department of Molecular and Developmental Genetics, Institute of Biology, Leiden University, Sylviusweg 72, 2333 BE, Leiden, The Netherlands

    Qi Jia, Amke den Dulk-Ras, Hexi Shen, Paul J. J. Hooykaas & Sylvia de Pater

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  2. Amke den Dulk-Ras
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Correspondence to Sylvia de Pater.

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Jia, Q., Dulk-Ras, A.d., Shen, H. et al. Poly(ADP-ribose)polymerases are involved in microhomology mediated back-up non-homologous end joining in Arabidopsis thaliana . Plant Mol Biol 82, 339–351 (2013). https://doi.org/10.1007/s11103-013-0065-9

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