Heterochromatin loosening by the Oct4 linker region facilitates Klf4 binding and iPSC reprogramming

EMBO J. 2020 Jan 2;39(1):e99165. doi: 10.15252/embj.201899165. Epub 2019 Oct 1.

Abstract

The success of Yamanaka factor reprogramming of somatic cells into induced pluripotent stem cells suggests that some factor(s) must remodel the nuclei from a condensed state to a relaxed state. How factor-dependent chromatin opening occurs remains unclear. Using FRAP and ATAC-seq, we found that Oct4 acts as a pioneer factor that loosens heterochromatin and facilitates the binding of Klf4 and the expression of epithelial genes in early reprogramming, leading to enhanced mesenchymal-to-epithelial transition. A mutation in the Oct4 linker, L80A, which shows impaired interaction with the BAF complex component Brg1, is inactive in heterochromatin loosening. Oct4-L80A also blocks the binding of Klf4 and retards MET. Finally, vitamin C or Gadd45a could rescue the reprogramming deficiency of Oct4-L80A by enhancing chromatin opening and Klf4 binding. These studies reveal a cooperation between Oct4 and Klf4 at the chromatin level that facilitates MET at the cellular level and shed light into the research of multiple factors in cell fate determination.

Keywords: Klf4; Oct4; heterochromatin loosening; mesenchymal-to-epithelial transition; reprogramming.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antioxidants / pharmacology
  • Ascorbic Acid / pharmacology
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism
  • Cell Differentiation
  • Cells, Cultured
  • Cellular Reprogramming*
  • DNA Helicases / genetics
  • DNA Helicases / metabolism
  • Epithelial Cells / cytology
  • Epithelial Cells / metabolism*
  • Epithelial-Mesenchymal Transition
  • Fibroblasts / cytology
  • Fibroblasts / metabolism
  • Heterochromatin / genetics
  • Heterochromatin / metabolism*
  • Histones / genetics
  • Histones / metabolism*
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Kruppel-Like Factor 4
  • Kruppel-Like Transcription Factors / genetics
  • Kruppel-Like Transcription Factors / metabolism*
  • Mice
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Octamer Transcription Factor-3 / genetics
  • Octamer Transcription Factor-3 / metabolism*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism

Substances

  • Antioxidants
  • Cell Cycle Proteins
  • GADD45A protein, human
  • Heterochromatin
  • Histones
  • KLF4 protein, human
  • Klf4 protein, mouse
  • Kruppel-Like Factor 4
  • Kruppel-Like Transcription Factors
  • Nuclear Proteins
  • Octamer Transcription Factor-3
  • POU5F1 protein, human
  • Transcription Factors
  • histone H3 trimethyl Lys4
  • SMARCA4 protein, human
  • DNA Helicases
  • Ascorbic Acid

Associated data

  • GEO/GSE129728

Grants and funding