Dynamic Reorganization of Nucleosome Positioning in Somatic Cells after Transfer into Porcine Enucleated Oocytes

Stem Cell Reports. 2017 Aug 8;9(2):642-653. doi: 10.1016/j.stemcr.2017.06.004. Epub 2017 Jul 6.

Abstract

The nucleosome, the fundamental structural unit of chromatin, is a critical regulator of gene expression. The mechanisms governing changes to nucleosome occupancy and positioning during somatic cell reprogramming remain poorly understood. We established a method for generating genome-wide nucleosome maps of porcine embryonic fibroblasts (PEF), reconstructed 1-cell embryos generated by somatic cell nuclear transfer (SCNT), and fertilized zygotes (FZ) using MNase sequencing with only 1,000 cells. We found that donor PEF chromatin, especially X chromosome, became more open after transfer into porcine oocytes and nucleosome occupancy decreased in promoters but increased in the genic regions. Nucleosome arrangements around transcriptional start sites of genes with different expression levels in somatic cells tended to become transcriptionally silent in SCNT; however, some pluripotency genes adopted transcriptionally active nucleosome arrangements. FZ and SCNT had similar characteristics, unlike PEF. This study reveals the dynamics and importance of nucleosome positioning and chromatin organization early after reprogramming.

Keywords: FZ; PEF; SCNT; gene expression; handmade cloning; nucleosome organization; reprogramming.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cellular Reprogramming*
  • Chromatin Assembly and Disassembly
  • Female
  • Fertilization in Vitro
  • Gene Expression
  • Male
  • Nuclear Transfer Techniques*
  • Nucleosomes / metabolism*
  • Oocytes / cytology*
  • Oocytes / metabolism*
  • Protein Binding
  • Sequence Analysis, DNA
  • Swine
  • Transcription Initiation Site
  • Transcriptional Activation

Substances

  • Nucleosomes