O-GlcNAc regulates pluripotency and reprogramming by directly acting on core components of the pluripotency network

Cell Stem Cell. 2012 Jul 6;11(1):62-74. doi: 10.1016/j.stem.2012.03.001. Epub 2012 May 17.

Abstract

O-linked-N-acetylglucosamine (O-GlcNAc) has emerged as a critical regulator of diverse cellular processes, but its role in embryonic stem cells (ESCs) and pluripotency has not been investigated. Here we show that O-GlcNAcylation directly regulates core components of the pluripotency network. Blocking O-GlcNAcylation disrupts ESC self-renewal and reprogramming of somatic cells to induced pluripotent stem cells. The core reprogramming factors Oct4 and Sox2 are O-GlcNAcylated in ESCs, but the O-GlcNAc modification is rapidly removed upon differentiation. O-GlcNAc modification of threonine 228 in Oct4 regulates Oct4 transcriptional activity and is important for inducing many pluripotency-related genes, including Klf2, Klf5, Nr5a2, Tbx3, and Tcl1. A T228A point mutation that eliminates this O-GlcNAc modification reduces the capacity of Oct4 to maintain ESC self-renewal and reprogram somatic cells. Overall, our study makes a direct connection between O-GlcNAcylation of key regulatory transcription factors and the activity of the pluripotency network.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylglucosamine / chemistry
  • Acetylglucosamine / pharmacology*
  • Amino Acid Sequence
  • Animals
  • Cell Differentiation / drug effects
  • Cell Differentiation / genetics
  • Cell Proliferation / drug effects
  • Cellular Reprogramming / drug effects*
  • Cellular Reprogramming / genetics
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / metabolism
  • Gene Expression Regulation, Developmental / drug effects
  • Gene Regulatory Networks / drug effects*
  • Glycosylation / drug effects
  • Humans
  • Mice
  • Mice, Inbred C57BL
  • Models, Biological
  • Molecular Sequence Data
  • Mutation / genetics
  • Octamer Transcription Factor-3 / chemistry
  • Octamer Transcription Factor-3 / genetics
  • Octamer Transcription Factor-3 / metabolism
  • Pluripotent Stem Cells / drug effects
  • Pluripotent Stem Cells / metabolism*
  • SOXB1 Transcription Factors / metabolism
  • Transcription, Genetic / drug effects

Substances

  • Octamer Transcription Factor-3
  • Pou5f1 protein, mouse
  • SOXB1 Transcription Factors
  • Sox2 protein, mouse
  • Acetylglucosamine