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Arabidopsis DEMETER-LIKE proteins DML2 and DML3 are required for appropriate distribution of DNA methylation marks

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Abstract

Cytosine DNA methylation is a stable epigenetic mark for maintenance of gene silencing across cellular divisions, but it is a reversible modification. Genetic and biochemical studies have revealed that the Arabidopsis DNA glycosylase domain-containing proteins ROS1 (REPRESSOR OF SILENCING 1) and DME (DEMETER) initiate erasure of 5-methylcytosine through a base excision repair process. The Arabidopsis genome encodes two paralogs of ROS1 and DME, referred to as DEMETER-LIKE proteins DML2 and DML3. We have found that DML2 and DML3 are 5-methylcytosine DNA glycosylases that are expressed in a wide range of plant organs. We analyzed the distribution of methylation marks at two methylated loci in wild-type and dml mutant plants. Mutations in DML2 and/or DML3 lead to hypermethylation of cytosine residues that are unmethylated or weakly methylated in wild-type plants. In contrast, sites that are heavily methylated in wild-type plants are hypomethylated in mutants. These results suggest that DML2 and DML3 are required not only for removing DNA methylation marks from improperly-methylated cytosines, but also for maintenance of high methylation levels in properly targeted sites.

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Abbreviations

5-meC:

5-Methylcytosine

DME :

DEMETER

DML :

DEMETER-LIKE

MBP:

Maltose-binding protein

ROS1 :

REPRESSOR OF SILENCING 1

TDG:

Thymine DNA glycosylase

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Acknowledgements

This research was supported by Ministerio de Educación y Ciencia (Grant BFU2004-05303) and Junta de Andalucía (CVI-301).

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Authors and Affiliations

  1. Departamento de Genética, Universidad de Córdoba, Edificio Gregor Mendel, Campus de Rabanales s/n, 14071, Córdoba, Spain

    Ana Pilar Ortega-Galisteo, Teresa Morales-Ruiz, Rafael R. Ariza & Teresa Roldán-Arjona

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  1. Ana Pilar Ortega-Galisteo
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  2. Teresa Morales-Ruiz
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  3. Rafael R. Ariza
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  4. Teresa Roldán-Arjona
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Correspondence to Teresa Roldán-Arjona.

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Ortega-Galisteo, A.P., Morales-Ruiz, T., Ariza, R.R. et al. Arabidopsis DEMETER-LIKE proteins DML2 and DML3 are required for appropriate distribution of DNA methylation marks. Plant Mol Biol 67, 671–681 (2008). https://doi.org/10.1007/s11103-008-9346-0

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