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PMID: 22325353 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

A DNA 3' phosphatase functions in active DNA demethylation in Arabidopsis.

Molecular cell ·Vol. 45 ·No. 3 ·2012-02-10 ·Pages 357-70

Martínez-Macías MI, Qian W, Miki D, Pontes O, Liu Y, Tang K, Liu R, Morales-Ruiz T, Ariza RR, Roldán-Arjona T, Zhu JK

Abstract

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 colocalize 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.

MeSH Terms
5-Methylcytosine/metabolism Arabidopsis/enzymology,genetics Arabidopsis Proteins/chemistry,metabolism Base Sequence Cell Nucleus/metabolism DNA Cleavage DNA Methylation Gene Expression Regulation, Plant Gene Silencing Genes, Reporter Genetic Loci Genome, Plant Kinetics Luciferases/biosynthesis,genetics Molecular Sequence Data Mutagenesis, Insertional Nuclear Proteins/chemistry,metabolism Nucleotidases/chemistry,genetics,metabolism Protein Binding RNA-Binding Proteins/metabolism Transcription, Genetic
Chemicals
Arabidopsis Proteins Nuclear Proteins RNA-Binding Proteins ROS1 protein, Arabidopsis ROS3 protein, Arabidopsis 5-Methylcytosine Luciferases Nucleotidases deoxynucleotide 3'-phosphatase
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Martínez-Macías María Isabel
Department of Genetics, University of Córdoba, 14071 Córdoba, Spain.
Qian Weiqiang
Miki Daisuke
Pontes Olga
Liu Yunhua
Tang Kai
Liu Renyi
Morales-Ruiz Teresa
Ariza Rafael R
Roldán-Arjona Teresa
Zhu Jian-Kang
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Article Info
Journal
Molecular cell
Abbr.
Mol Cell
ISSN
1097-4164
DOI
10.1016/j.molcel.2011.11.034
Published
2012-02-10
Pages
357-70
Language
English
Region
United States
NLM ID
9802571
PMCID
PMC3278721
Subset
IM
Grants
NIGMS NIH HHS · R01 GM070795 · United States
NIGMS NIH HHS · R01 GM070795-09 · United States
NIGMS NIH HHS · R01GM070795 · United States
External Links
PubMed source DOI: 10.1016/j.molcel.2011.11.034 PMC Full Text View journal details
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