Rapid repair of DNA double strand breaks in Arabidopsis thaliana is dependent on proteins involved in chromosome structure maintenance

Kozák, Jaroslav; West, Claudia; White, Charles I.; Costa-Nunes, José A. da; Angelis, Karel J.

doi:10.1016/j.dnarep.2008.11.012

Cited by 71 publications

(76 citation statements)

References 41 publications

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“…After a 60-min recovery period, 33% 6 11% of wild-type DNA still localized within the tail, while tail DNA levels were considerably higher in ctf7 (61% 6 9%), wapl1-1 wapl2 ctf7 (59% 6 8%), and wapl1-1 wapl2 (56% 6 7%) plants ( Figures 2B and 2C). Calculation of DNA repair efficiencies (Kozak et al, 2009) indicated that 20% 6 9% of DSB's remained unrepaired after a 60-min recovery period in wild-type plants, while 75% 6 9%, 76% 6 8%, and 80% 6 9% of DBS's remained unrepaired in ctf7, wapl1-1 wapl2, and wapl1-1 wapl2 ctf7 plants, respectively. Plants lacking PDS5 also show alterations in DNA repair (Pradillo et al, 2015), indicating that alterations in sister chromatid cohesion in general affect DNA repair rates.…”

Section: Inactivation Of Wapl Fails To Rescue Ctf7-associated Somaticmentioning

confidence: 99%

“…At least 300 nuclei per line were analyzed. Damage remaining after a given repair time (tx) was estimated according to Kozak et al (2009). Results were obtained from at least three different biological samples, with three technical repeats and a negative control for each line.…”

Section: Flow Cytometrymentioning

confidence: 99%

“…Short treatments (1 to 2 h.) with Mitomycin C have been shown to be effective in generating DNA DSBs (Bohmdorfer et al, 2011). After incubation for 2 h in the dark, the plants were washed three times with distilled water and incubated for 1 h in liquid 0.53 MS medium without Mitomycin C. Leaves were excised, prepared, and then processed with a Trevigen CometAssay Kit (Trevigen) according to the manufacturer's instructions and published procedures (Collins et al, 2008;Kozak et al, 2009). Peroxide was used as a positive control for the oxidation status of the cells (Collins, 2014).…”

Section: Flow Cytometrymentioning

confidence: 99%

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The Opposing Actions of Arabidopsis CHROMOSOME TRANSMISSION FIDELITY7 and WINGS APART-LIKE1 and 2 Differ in Mitotic and Meiotic Cells

Bolaños-Villegas

Mitra

et al. 2016

Plant Cell

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Sister chromatid cohesion, which is mediated by the cohesin complex, is essential for the proper segregation of chromosomes during mitosis and meiosis. Stable binding of cohesin with chromosomes is regulated in part by the opposing actions of CTF7 (CHROMOSOME TRANSMISSION FIDELITY7) and WAPL (WINGS APART-LIKE). In this study, we characterized the interaction between Arabidopsis thaliana CTF7 and WAPL by conducting a detailed analysis of wapl1-1 wapl2 ctf7 plants. ctf7 plants exhibit major defects in vegetative growth and development and are completely sterile. Inactivation of WAPL restores normal growth, mitosis, and some fertility to ctf7 plants. This shows that the CTF7/WAPL cohesin system is not essential for mitosis in vegetative cells and suggests that plants may contain a second mechanism to regulate mitotic cohesin. WAPL inactivation restores cohesin binding and suppresses ctf7-associated meiotic cohesion defects, demonstrating that WAPL and CTF7 function as antagonists to regulate meiotic sister chromatid cohesion. The ctf7 mutation only had a minor effect on wapl-associated defects in chromosome condensation and centromere association. These results demonstrate that WAPL has additional roles that are independent of its role in regulating chromatin-bound cohesin.

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Section: Inactivation Of Wapl Fails To Rescue Ctf7-associated Somaticmentioning

confidence: 99%

Section: Flow Cytometrymentioning

confidence: 99%

Section: Flow Cytometrymentioning

confidence: 99%

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The Opposing Actions of Arabidopsis CHROMOSOME TRANSMISSION FIDELITY7 and WINGS APART-LIKE1 and 2 Differ in Mitotic and Meiotic Cells

Bolaños-Villegas

Mitra

et al. 2016

Plant Cell

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“…The extent of the accumulation of nuclear DNA damage in the wild type and mutant lines in response to high salinity and other chemical agents was analyzed by comet assay under neutral conditions to detect the DSBs as described previously Kozak et al (2009).…”

Section: Analysis Of the Induction And Repair Of Dsbs By Comet Assaymentioning

confidence: 99%

“…Since high salinity is known to induce DSBs in DNA and Pol l has been implicated in NHEJ-mediated DSB repair (Capp et al, 2006;Moon et al, 2007;Garcia-Diaz et al, 2009), we next analyzed the extent of DSB accumulation and repair in the wild type and atpoll mutants following the exposure of seedlings to high salinity (200 mM NaCl). A comet assay was carried out under neutral conditions (Kozak et al, 2009) using a nuclear suspension prepared from control and salt-treated 7-d-old seedlings to measure the relative accumulation and repair of DSBs (Supplemental Fig. S9, A-F).…”

Section: Abiotic and Genotoxic Stress Induces Atpoll Expressionmentioning

confidence: 99%

Involvement of AtPolλ in the Repair of High Salt- and DNA Cross-Linking Agent-Induced Double Strand Breaks in Arabidopsis

et al. 2013

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DNA polymerase l (Pol l) is the sole member of family X DNA polymerase in plants and plays a crucial role in nuclear DNA damage repair. Here, we report the transcriptional up-regulation of Arabidopsis (Arabidopsis thaliana) AtPoll in response to abiotic and genotoxic stress, including salinity and the DNA cross-linking agent mitomycin C (MMC). The increased sensitivity of atpoll knockout mutants toward high salinity and MMC treatments, with higher levels of accumulation of double strand breaks (DSBs) than wild-type plants and delayed repair of DSBs, has suggested the requirement of Pol l in DSB repair in plants.AtPoll overexpression moderately complemented the deficiency of DSB repair capacity in atpoll mutants. Transcriptional upregulation of major nonhomologous end joining (NHEJ) pathway genes KU80, X-RAY CROSS COMPLEMENTATION PROTEIN4 (XRCC4), and DNA Ligase4 (Lig4) along with AtPoll in Arabidopsis seedlings, and the increased sensitivity of atpoll-2/atxrcc4 and atpoll-2/atlig4 double mutants toward high salinity and MMC treatments, indicated the involvement of NHEJ-mediated repair of salinity-and MMC-induced DSBs. The suppressed expression of NHEJ genes in atpoll mutants suggested complex transcriptional regulation of NHEJ genes. Pol l interacted directly with XRCC4 and Lig4 via its N-terminal breast cancer-associated C terminus (BRCT) domain in a yeast two-hybrid system, while increased sensitivity of BRCT-deficient Pol l-expressing transgenic atpoll-2 mutants toward genotoxins indicated the importance of the BRCT domain of AtPoll in mediating the interactions for processing DSBs. Our findings provide evidence for the direct involvement of DNA Pol l in the repair of DSBs in a plant genome.

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Telomeres in Plant Meiosis: Their Structure, Dynamics and Function

Roberts

Osman

Chris

et al. 2013

Annual Plant Reviews

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Rapid repair of DNA double strand breaks in Arabidopsis thaliana is dependent on proteins involved in chromosome structure maintenance

Cited by 71 publications

References 41 publications

The Opposing Actions of Arabidopsis CHROMOSOME TRANSMISSION FIDELITY7 and WINGS APART-LIKE1 and 2 Differ in Mitotic and Meiotic Cells

The Opposing Actions of Arabidopsis CHROMOSOME TRANSMISSION FIDELITY7 and WINGS APART-LIKE1 and 2 Differ in Mitotic and Meiotic Cells

Involvement of AtPolλ in the Repair of High Salt- and DNA Cross-Linking Agent-Induced Double Strand Breaks in Arabidopsis

Telomeres in Plant Meiosis: Their Structure, Dynamics and Function

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