Hierarchical molecular events driven by oocyte-specific factors lead to rapid and extensive reprogramming

Mol Cell. 2014 Aug 21;55(4):524-36. doi: 10.1016/j.molcel.2014.06.024. Epub 2014 Jul 24.

Abstract

Nuclear transfer to oocytes is an efficient way to transcriptionally reprogram somatic nuclei, but its mechanisms remain unclear. Here, we identify a sequence of molecular events that leads to rapid transcriptional reprogramming of somatic nuclei after transplantation to Xenopus oocytes. RNA-seq analyses reveal that reprogramming by oocytes results in a selective switch in transcription toward an oocyte rather than pluripotent type, without requiring new protein synthesis. Time-course analyses at the single-nucleus level show that transcriptional reprogramming is induced in most transplanted nuclei in a highly hierarchical manner. We demonstrate that an extensive exchange of somatic- for oocyte-specific factors mediates reprogramming and leads to robust oocyte RNA polymerase II binding and phosphorylation on transplanted chromatin. Moreover, genome-wide binding of oocyte-specific linker histone B4 supports its role in transcriptional reprogramming. Thus, our study reveals the rapid, abundant, and stepwise loading of oocyte-specific factors onto somatic chromatin as important determinants for successful reprogramming.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cells, Cultured
  • Cellular Reprogramming / genetics*
  • Cellular Reprogramming / physiology
  • Chromatin / metabolism*
  • Genome
  • Histones / physiology*
  • Mice
  • Nuclear Transfer Techniques
  • Oocytes / metabolism*
  • Organ Specificity
  • RNA / genetics
  • Sequence Analysis, RNA
  • Xenopus / embryology*
  • Xenopus / genetics

Substances

  • Chromatin
  • Histones
  • RNA

Associated data

  • SRA/SRP042359