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Nucleolar Reorganization Upon Site-Specific Double-Strand Break Induction.

Michal Franek1, Alena Kovaříková1, Eva Bártová1

  • 1Institute of Biophysics, Academy of Sciences of the Czech Republic, Brno, Czech Republic (MF, AK, EB, SK).

The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
|September 30, 2016
PubMed
Summary

Embryonic stem cells exhibit unique DNA damage responses in nucleoli, forming protein-rich caps after ribosomal gene breaks. These caps involve specific proteins and modifications, but not heterochromatin protein 1.

Keywords:
CARM1DNA repairHDAC1NBS1PpoIchromatinnucleolus

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • DNA damage response (DDR) in embryonic stem cells (ESCs) is unique due to their rapid proliferation and specific chromatin.
  • The nucleolus, crucial for rRNA production in ESCs, has poorly understood DDR mechanisms.
  • Ribosomal gene transcription in the nucleolus is vital but its DNA repair pathways remain largely unexplored.

Purpose of the Study:

  • To investigate DNA damage response and repair mechanisms within the nucleolus of ESCs.
  • To characterize the formation and protein composition of nucleolar caps induced by DNA double-strand breaks in rRNA genes.

Main Methods:

  • Induced double-strand breaks in rRNA genes using I-PpoI endonuclease in ESCs.
  • Analyzed nucleolar morphology, DDR protein recruitment (BRCA1, NBS1, MDC1, γH2AX, UBF1), and chromatin modifications.
  • Investigated interactions of nucleolar proteins (TCOF1) with enzymes (HDAC1, CARM1) and histone modifications (H3R17me2a).

Main Results:

  • Observed significant formation of I-PpoI-induced nucleolar caps containing key DDR proteins.
  • Demonstrated interactions between TCOF1, HDAC1, and CARM1 following DNA injury.
  • Identified H3R17me2a modification within nucleolar caps, mediated by CARM1.
  • Reported that heterochromatin protein 1 is not involved in the repair of these nucleolar caps.

Conclusions:

  • ESCs mount a distinct DDR in the nucleolus, characterized by nucleolar cap formation.
  • Specific protein interactions and histone modifications are crucial for nucleolar DNA repair in ESCs.
  • The study elucidates novel aspects of DNA repair within the nucleolus of pluripotent stem cells.