<i>Ku80‐</i> and <i>DNA ligase IV</i>‐deficient plants are sensitive to ionizing radiation and defective in T‐DNA integration

Friesner, Joanna; Britt, Anne B.

doi:10.1046/j.1365-313x.2003.01738.x

Cited by 152 publications

(150 citation statements)

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“…There are conflicting reports about the role of KU80 in T-DNA integration. Gallego et al (2003) found no deficiency in T-DNA integration in Arabidopsis ku80 mutants, whereas Friesner and Britt (2003) showed a reduction in T-DNA integration in Arabidopsis ku80 mutants. Subsequently, Li et al (2005) showed that Arabidopsis ku80 mutants showed decreased stable transformation, whereas Ku80 overexpression showed increased T-DNA integration and increased resistance to methyl methanesulfonate (MMS).…”

Section: Introductionmentioning

confidence: 89%

Agrobacterium May Delay Plant Nonhomologous End-Joining DNA Repair via XRCC4 to Favor T-DNA Integration

et al. 2012

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Agrobacterium tumefaciens is a soilborne pathogen that causes crown gall disease in many dicotyledonous plants by transfer of a portion of its tumor-inducing plasmid (T-DNA) into the plant genome. Several plant factors that play a role in Agrobacterium attachment to plant cells and transport of T-DNA to the nucleus have been identified, but the T-DNA integration step during transformation is poorly understood and has been proposed to occur via nonhomologous end-joining (NHEJ)-mediated doublestrand DNA break (DSB) repair. Here, we report a negative role of X-RAY CROSS COMPLEMENTATION GROUP4 (XRCC4), one of the key proteins required for NHEJ, in Agrobacterium T-DNA integration. Downregulation of XRCC4 in Arabidopsis and Nicotiana benthamiana increased stable transformation due to increased T-DNA integration. Overexpression of XRCC4 in Arabidopsis decreased stable transformation due to decreased T-DNA integration. Interestingly, XRCC4 directly interacted with Agrobacterium protein VirE2 in a yeast two-hybrid system and in planta. VirE2-expressing Arabidopsis plants were more susceptible to the DNA damaging chemical bleomycin and showed increased stable transformation. We hypothesize that VirE2 titrates or excludes active XRCC4 protein available for DSB repair, thus delaying the closure of DSBs in the chromosome, providing greater opportunity for T-DNA to integrate.

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Section: Introductionmentioning

confidence: 89%

Agrobacterium May Delay Plant Nonhomologous End-Joining DNA Repair via XRCC4 to Favor T-DNA Integration

et al. 2012

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“…It has been shown recently that these difficulties may be caused by the existence of independent end-joining pathways in plant cells (Charbonnel et al, 2011). A mild reduction in T-DNA transformation efficiencies was reported when single NHEJ factors were knocked out, but no GT experiments have yet been published concerning this issue (Friesner and Britt, 2003;Nishizawa-Yokoi et al, 2012).…”

Section: Manipulation Of the Enzyme Machinery Can Help (A Bit)mentioning

confidence: 99%

Gene targeting in plants: 25 years later

Puchta

Fauser²

2013

Int. J. Dev. Biol.

164

115

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Only five years after the initiation of transgenic research in plants, gene targeting (GT) was achieved for the first time in tobacco. Unfortunately, the frequency of targeted integration via homologous recombination (HR) was so low in comparison to random integration that GT could not be established as a feasible technique in higher plants. It took another 25 years and great effort to develop the knowledge and tools necessary to overcome this challenge, at least for some plant species. In some cases, the overexpression of proteins involved in HR or the use of negative selectable markers improved GT to a certain extent. An effective solution to this problem was developed in 1996, when a sequence-specific endonuclease was used to induce a double-strand break (DSB) at the target locus. Thus, GT frequencies were enhanced dramatically. Thereafter, the main limitation was the absence of tools needed to induce DSBs at specific sites in the genome. Such tools became available with the development of zinc finger nucleases (ZFNs), and a breakthrough was achieved in 2005 when ZFNs were used to target a marker gene in tobacco. Subsequently, endogenous loci were targeted in maize, tobacco and Arabidopsis. Recently, our toolbox for genetic engineering has expanded with the addition of more types of site-specific endonucleases, meganucleases, transcription activator-like effector nucleases (TALENs) and the CRISPR/Cas system. We assume that targeted genome modifications will become routine in the near future in crop plants using these nucleases along with the newly developed in planta GT technique.

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“…Mutants in DNA repair/maintenance genes in Arabidopsis, such as DNA ligase IV, ERCC1, and telomerase, do not exhibit obvious cell death, even in response to treatment with DNA damaging agents (Riha et al, 2001;Friesner and Britt, 2003;Hefner et al, 2003). A prevailing hypothesis is that plants, which have no p53 homolog, lack DNA damage-induced cell death (apoptosis).…”

Section: Atatr Responses To Replication Blocksmentioning

confidence: 99%

ATR Regulates a G2-Phase Cell-Cycle Checkpoint in Arabidopsis thaliana

2004

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Ataxia telangiectasia-mutated and Rad3-related (ATR) plays a central role in cell-cycle regulation, transmitting DNA damage signals to downstream effectors of cell-cycle progression. In animals, ATR is an essential gene. Here, we find that Arabidopsis (Arabidopsis thaliana) atrÿ/ÿ mutants were viable, fertile, and phenotypically wild-type in the absence of exogenous DNA damaging agents but exhibit altered expression of AtRNR1 (ribonucleotide reductase large subunit) and alteration of some damage-induced cell-cycle checkpoints. atr mutants were hypersensitive to hydroxyurea (HU), aphidicolin, and UV-B light but only mildly sensitive to g-radiation. G2 arrest was observed in response to g-irradiation in both wild-type and atr plants, albeit with slightly different kinetics, suggesting that ATR plays a secondary role in response to double-strand breaks. G2 arrest also was observed in wild-type plants in response to aphidicolin but was defective in atr mutants, resulting in compaction of nuclei and subsequent cell death. By contrast, HU-treated wild-type and atr plants arrested in G1 and showed no obvious signs of cell death. We propose that, in plants, HU invokes a novel checkpoint responsive to low levels of deoxynucleotide triphosphates. These results demonstrate the important role of cell-cycle checkpoints in the ability of plant cells to sense and cope with problems associated with DNA replication.

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Ku80‐ and DNA ligase IV‐deficient plants are sensitive to ionizing radiation and defective in T‐DNA integration

Cited by 152 publications

References 50 publications

Agrobacterium May Delay Plant Nonhomologous End-Joining DNA Repair via XRCC4 to Favor T-DNA Integration

Agrobacterium May Delay Plant Nonhomologous End-Joining DNA Repair via XRCC4 to Favor T-DNA Integration

Gene targeting in plants: 25 years later

ATR Regulates a G2-Phase Cell-Cycle Checkpoint in Arabidopsis thaliana

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