Arabidopsis Uracil DNA Glycosylase (UNG) Is Required for Base Excision Repair of Uracil and Increases Plant Sensitivity to 5-Fluorouracil

Córdoba-Cañero, Dolores; Dubois, Emeline; Ariza, Rafael R.; Doutriaux, Marie-Pascale; Roldán-Arjona, Teresa

doi:10.1074/jbc.m109.067173

Cited by 43 publications

(56 citation statements)

References 37 publications

(39 reference statements)

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“…, 2009). Consistent with our previously reported observations (Córdoba‐Cañero et al. , 2009, 2010), WT Arabidopsis extracts are capable of performing full BER on both substrates in a reaction that is dependent on the presence of deoxyribonucleoside triphosphates (dNTPs) in the mixture (Figure 1c, lanes 3 and 8).…”

Section: Resultssupporting

confidence: 91%

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Arabidopsis ARP endonuclease functions in a branched base excision DNA repair pathway completed by LIG1

Córdoba-Cañero

Roldán-Arjona

Ariza³

2011

The Plant Journal

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SUMMARYBase excision repair (BER) is an essential cellular defence mechanism against DNA damage, but it is poorly understood in plants. We used an assay that monitors repair of damaged bases and abasic (apurinic/ apyrimidinic, AP) sites in Arabidopsis to characterize post-excision events during plant BER. We found that Apurinic endonuclease-redox protein (ARP) is the major AP endonuclease activity in Arabidopsis cell extracts, and is required for AP incision during uracil BER in vitro. Mutant plants that are deficient in ARP grow normally but are hypersensitive to 5-fluorouracil, a compound that favours mis-incorporation of uracil into DNA. We also found that, after AP incision, the choice between single-nucleotide or long-patch DNA synthesis (SN-or LP-BER) is influenced by the 5¢ end of the repair gap. When the 5¢ end is blocked and not amenable to b-elimination, the SN sub-pathway is abrogated, and repair is accomplished through LP-BER only. Finally, we provide evidence that Arabidopsis DNA ligase I (LIG1) is required for both SN-and LP-BER. lig1 RNAi-silenced lines show very reduced uracil BER, and anti-LIG1 antibody abolishes repair in wild-type cell extracts. In contrast, knockout lig4 )/) mutants exhibit normal BER and nick ligation levels. Our results suggest that a branched BER pathway completed by a member of the DNA ligase I family may be an ancient feature in eukaryotic species.

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Section: Resultssupporting

confidence: 91%

“…, 2009). Using this assay, we found that uracil repair in Arabidopsis is initiated by the uracil DNA glycosylase AtUNG (Córdoba‐Cañero et al. , 2010), and that the ensuing DNA repair intermediates are processed by both AP lyases and AP endonucleases (Córdoba‐Cañero et al.…”

Section: Introductionmentioning

confidence: 99%

Arabidopsis ARP endonuclease functions in a branched base excision DNA repair pathway completed by LIG1

Córdoba-Cañero

Roldán-Arjona

Ariza³

2011

The Plant Journal

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“…We have found that ZDP is the major, if not the only, DNA 3′-phosphatase activity in Arabidopsis whole cell extracts, and it is strictly required for complete processing of part of the DNA demethylation intermediates generated by ROS1. An additional important finding of this work is that, after ZDP-dependent 3′-end cleaning, replacement of the excised 5-meC by an unmethylated cytosine may be carried out both via single-nucleotide insertion and long-patch DNA synthesis, as previously reported in Arabidopsis for other DNA modifications (Córdoba-Cañero et al, 2010; Córdoba-Cañero et al, 2009). The existence of two different sub-pathways for gap-filling and ligation expands the spectrum of potential proteins, particularly DNA polymerases, that may participate in the post-incision events that take place during BER demethylation.…”

Section: Discussionmentioning

confidence: 53%

A DNA 3′ Phosphatase Functions in Active DNA Demethylation in Arabidopsis

et al. 2012

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SUMMARY DNA methylation is an important epigenetic mark established by the combined actions of methylation and demethylation reactions. Plants use a base excision repair pathway for active DNA demethylation. After 5-methylcytosine removal, the Arabidopsis DNA glycosylase/lyase ROS1 incises the DNA backbone and part of the product has a single-nucleotide gap flanked by 3′- and 5′-phosphate termini. Here we show that the DNA phosphatase ZDP removes the blocking 3′-phosphate, allowing subsequent DNA polymerization and ligation steps needed to complete the repair reactions. ZDP and ROS1 interact in vitro and co-localize in vivo in nucleoplasmic foci. Extracts from zdp mutant plants are unable to complete DNA demethylation in vitro, and the mutations cause DNA hypermethylation and transcriptional silencing of a reporter gene. Genome-wide methylation analysis in zdp mutant plants identified hundreds of hypermethylated endogenous loci. Our results show that ZDP functions downstream of ROS1 in one branch of the active DNA demethylation pathway.

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“…Maximum likelihood tree obtained using the RAxML tool with 100 bootstrap replicates. Gray clades grouped protein sequences closer to proteins of known function: AlkA: alkyladenine glycosylases that excise 3‐methyladenine (Begley et al, ); Nth: endonuclease III and OggI: 8‐oxoguanine glycosylase I, which remove oxidative damaged bases from the DNA backbone (Rosenquist, Zharkov, & Grollman, ); U: specific for the excision of uracil (Córdoba‐Cañero, Dubois, Ariza, Doutriaux, & Roldán‐Arjona, ); mA: excise 3‐methyl adenine (Santerre & Britt, ); MutM: formamidopyrimidine‐DNA glycosylase (Duclos et al, ); 5mC: the group of 5mC DNA glycosylase (Zhu, ). CsDML and five poplar DML proteins clustered with AtDME, AtROS1, AtDML2 and AtDML3.…”

Section: Resultsmentioning

confidence: 99%

Overexpression of DEMETER, a DNA demethylase, promotes early apical bud maturation in poplar

Conde

Moreno-Cortés

Dervinis

et al. 2017

Plant Cell & Environment

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The transition from active growth to dormancy is critical for the survival of perennial plants. We identified a DEMETER-like (CsDML) cDNA from a winter-enriched cDNA subtractive library in chestnut (Castanea sativa Mill.), an economically and ecologically important species. Next, we characterized this DNA demethylase and its putative ortholog in the more experimentally tractable hybrid poplar (Populus tremula × alba), under the signals that trigger bud dormancy in trees. We performed phylogenetic and protein sequence analysis, gene expression profiling, and 5-methyl-cytosine methylation immunodetection studies to evaluate the role of CsDML and its homolog in poplar, PtaDML6. Transgenic hybrid poplars overexpressing CsDML were produced and analysed. Short days and cold temperatures induced CsDML and PtaDML6. Overexpression of CsDML accelerated short-day-induced bud formation, specifically from Stages 1 to 0. Buds acquired a red-brown coloration earlier than wild-type plants, alongside with the up-regulation of flavonoid biosynthesis enzymes and accumulation of flavonoids in the shoot apical meristem and bud scales. Our data show that the CsDML gene induces bud formation needed for the survival of the apical meristem under the harsh conditions of winter.

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Arabidopsis Uracil DNA Glycosylase (UNG) Is Required for Base Excision Repair of Uracil and Increases Plant Sensitivity to 5-Fluorouracil

Cited by 43 publications

References 37 publications

Arabidopsis ARP endonuclease functions in a branched base excision DNA repair pathway completed by LIG1

Arabidopsis ARP endonuclease functions in a branched base excision DNA repair pathway completed by LIG1

A DNA 3′ Phosphatase Functions in Active DNA Demethylation in Arabidopsis

Overexpression of DEMETER, a DNA demethylase, promotes early apical bud maturation in poplar

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