A unique Oct4 interface is crucial for reprogramming to pluripotency

Nat Cell Biol. 2013 Mar;15(3):295-301. doi: 10.1038/ncb2680. Epub 2013 Feb 3.

Abstract

Terminally differentiated cells can be reprogrammed to pluripotency by the forced expression of Oct4, Sox2, Klf4 and c-Myc. However, it remains unknown how this leads to the multitude of epigenetic changes observed during the reprogramming process. Interestingly, Oct4 is the only factor that cannot be replaced by other members of the same family to induce pluripotency. To understand the unique role of Oct4 in reprogramming, we determined the structure of its POU domain bound to DNA. We show that the linker between the two DNA-binding domains is structured as an α-helix and exposed to the protein's surface, in contrast to the unstructured linker of Oct1. Point mutations in this α-helix alter or abolish the reprogramming activity of Oct4, but do not affect its other fundamental properties. On the basis of mass spectrometry studies of the interactome of wild-type and mutant Oct4, we propose that the linker functions as a protein-protein interaction interface and plays a crucial role during reprogramming by recruiting key epigenetic players to Oct4 target genes. Thus, we provide molecular insights to explain how Oct4 contributes to the reprogramming process.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Blotting, Western
  • Cell Differentiation*
  • Cells, Cultured
  • Cellular Reprogramming*
  • Crystallography, X-Ray
  • DNA / genetics
  • DNA / metabolism*
  • Electrophoretic Mobility Shift Assay
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / metabolism
  • Epigenesis, Genetic
  • Fibroblasts / cytology*
  • Fibroblasts / metabolism
  • Humans
  • Kruppel-Like Factor 4
  • Luciferases / metabolism
  • Mice
  • Molecular Sequence Data
  • Octamer Transcription Factor-3 / chemistry*
  • Octamer Transcription Factor-3 / genetics
  • Octamer Transcription Factor-3 / metabolism
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / metabolism
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Homology, Amino Acid
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • KLF4 protein, human
  • Klf4 protein, mouse
  • Kruppel-Like Factor 4
  • Octamer Transcription Factor-3
  • Pou5f1 protein, mouse
  • RNA, Messenger
  • DNA
  • Luciferases

Associated data

  • PDB/3L1P