title: Live-cell analysis of DNA methylation during sexual reproduction in Arabidopsis reveals context and sex-specific dynamics controlled by noncanonical RdDM.
abstract: Cytosine methylation is a key epigenetic mark in many organisms, important for both transcriptional control and genome integrity. While relatively stable during somatic growth, DNA methylation is reprogrammed genome-wide during mammalian reproduction. Reprogramming is essential for zygotic totipotency and to prevent transgenerational inheritance of epimutations. However, the extent of DNA methylation reprogramming in plants remains unclear. Here, we developed sensors reporting with single-cell resolution CG and non-CG methylation in Arabidopsis. Live imaging during reproduction revealed distinct and sex-specific dynamics for both contexts. We found that CHH methylation in the egg cell depends on DOMAINS REARRANGED METHYLASE 2 (DRM2) and RNA polymerase V (Pol V), two main actors of RNA-directed DNA methylation, but does not depend on Pol IV. Our sensors provide insight into global DNA methylation dynamics at the single-cell level with high temporal resolution and offer a powerful tool to track CG and non-CG methylation both during development and in response to environmental cues in all organisms with methylated DNA, as we illustrate in mouse embryonic stem cells.
authors.affiliation: Epigenetic Regulations and Seed Development, UMR232, Institut de Recherche pour le Développement (IRD), Université de Montpellier, 34394 Montpellier, France.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Ingouff
authors.affiliation: Epigenetic Regulations and Seed Development, UMR232, Institut de Recherche pour le Développement (IRD), Université de Montpellier, 34394 Montpellier, France.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Selles
authors.affiliation: Epigenetic Regulations and Seed Development, UMR232, Institut de Recherche pour le Développement (IRD), Université de Montpellier, 34394 Montpellier, France.
authors.forename: Caroline
authors.initials: C
authors.lastname: Michaud
authors.affiliation: Gregor Mendel Institute, Austrian Academy of Sciences, Vienna BioCenter, 1030 Vienna, Austria.
authors.forename: Thiet M
authors.initials: TM
authors.lastname: Vu
authors.affiliation: Gregor Mendel Institute, Austrian Academy of Sciences, Vienna BioCenter, 1030 Vienna, Austria.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Berger
authors.affiliation: Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724, USA.
authors.forename: Andrea J
authors.initials: AJ
authors.lastname: Schorn
authors.affiliation: Epigenetic Regulations and Seed Development, UMR232, Institut de Recherche pour le Développement (IRD), Université de Montpellier, 34394 Montpellier, France.
authors.forename: Daphné
authors.initials: D
authors.lastname: Autran
authors.affiliation: Department of Plant Systems Biology, VIB, Ghent University, B-9052 Ghent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium.
authors.forename: Matthias
authors.initials: M
authors.lastname: Van Durme
authors.affiliation: Department of Plant Systems Biology, VIB, Ghent University, B-9052 Ghent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium.
authors.forename: Moritz K
authors.initials: MK
authors.lastname: Nowack
authors.affiliation: Howard Hughes Medical Institute-Gordon and Betty Moore Foundation, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724, USA.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Martienssen
authors.affiliation: Epigenetic Regulations and Seed Development, UMR232, Institut de Recherche pour le Développement (IRD), Université de Montpellier, 34394 Montpellier, France.|Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Grimanelli
mesh_terms: D000818:Animals; D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000378:metabolism; D002460:Cell Line; D019175:DNA Methylation / Q000235:genetics*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D053595:Embryonic Stem Cells; D005786:Gene Expression Regulation; D008780:Methyltransferases / Q000235:genetics* / Q000378:metabolism*; D051379:Mice; D030821:Plants, Genetically Modified; D012098:Reproduction / Q000235:genetics; D012737:Sex Factors; D059010:Single-Cell Analysis
chemical_list: D029681:Arabidopsis Proteins; D008780:Methyltransferases; C470732:DRM2 protein, Arabidopsis; C538841:RNA polymerase V, Arabidopsis; D012321:DNA-Directed RNA Polymerases
keywords: DNA methylation; reproduction; reprogramming; sensors