Tunable DNMT1 degradation reveals DNMT1/DNMT3B synergy in DNA methylation and genome organization

Andrea Scelfo*, Viviana Barra, Nezar Abdennur, George Spracklin, Florence Busato, Catalina Salinas-Luypaert, Elena Bonaiti, Guillaume Velasco, Frédéric Bonhomme, Anna Chipont, Andréa E Tijhuis, Diana C J Spierings, Coralie Guérin, Paola Arimondo, Claire Francastel, Floris Foijer, Jӧrg Tost, Leonid Mirny, Daniele Fachinetti*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

5 Citations (Scopus)
13 Downloads (Pure)

Abstract

DNA methylation (DNAme) is a key epigenetic mark that regulates critical biological processes maintaining overall genome stability. Given its pleiotropic function, studies of DNAme dynamics are crucial, but currently available tools to interfere with DNAme have limitations and major cytotoxic side effects. Here, we present cell models that allow inducible and reversible DNAme modulation through DNMT1 depletion. By dynamically assessing whole genome and locus-specific effects of induced passive demethylation through cell divisions, we reveal a cooperative activity between DNMT1 and DNMT3B, but not of DNMT3A, to maintain and control DNAme. We show that gradual loss of DNAme is accompanied by progressive and reversible changes in heterochromatin, compartmentalization, and peripheral localization. DNA methylation loss coincides with a gradual reduction of cell fitness due to G1 arrest, with minor levels of mitotic failure. Altogether, this system allows DNMTs and DNA methylation studies with fine temporal resolution, which may help to reveal the etiologic link between DNAme dysfunction and human disease.

Original languageEnglish
Article numbere202307026
Number of pages19
JournalThe Journal of Cell Biology
Volume223
Issue number4
DOIs
Publication statusPublished - 1-Apr-2024

Keywords

  • Humans
  • DNA Methylation
  • Cell Division
  • Epigenomics
  • Heterochromatin/genetics

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