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CTCF chromatin residence time controls three-dimensional genome organization, gene expression and DNA methylation in pluripotent cells

  • Widia Soochit
  • , Frank Sleutels
  • , Gregoire Stik
  • , Marek Bartkuhn
  • , Sreya Basu
  • , Silvia C. Hernandez
  • , Sarra Merzouk
  • , Enrique Vidal
  • , Ruben Boers
  • , Joachim Boers
  • , Michael van der Reijden
  • , Bart Geverts
  • , Wiggert A. van Cappellen
  • , Mirjam van den Hout
  • , Zeliha Ozgur
  • , Wilfred F.J. van IJcken
  • , Joost Gribnau
  • , Rainer Renkawitz
  • , Thomas Graf
  • , Adriaan Houtsmuller
  • Frank Grosveld, Ralph Stadhouders, Niels Galjart

Research output: Contribution to journalArticlepeer-review

Abstract

The 11 zinc finger (ZF) protein CTCF regulates topologically associating domain formation and transcription through selective binding to thousands of genomic sites. Here, we replaced endogenous CTCF in mouse embryonic stem cells with green-fluorescent-protein-tagged wild-type or mutant proteins lacking individual ZFs to identify additional determinants of CTCF positioning and function. While ZF1 and ZF8–ZF11 are not essential for cell survival, ZF8 deletion strikingly increases the DNA binding off-rate of mutant CTCF, resulting in reduced CTCF chromatin residence time. Loss of ZF8 results in widespread weakening of topologically associating domains, aberrant gene expression and increased genome-wide DNA methylation. Thus, important chromatin-templated processes rely on accurate CTCF chromatin residence time, which we propose depends on local sequence and chromatin context as well as global CTCF protein concentration.

Original languageEnglish (US)
Pages (from-to)881-893
Number of pages13
JournalNature Cell Biology
Volume23
Issue number8
DOIs
StatePublished - Aug 2021
Externally publishedYes

ASJC Scopus subject areas

  • Cell Biology

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