Related Experiment Video

Updated: Jan 3, 2026

Investigation of Protein Recruitment to DNA Lesions Using 405 Nm Laser Micro-irradiation
12:29

Investigation of Protein Recruitment to DNA Lesions Using 405 Nm Laser Micro-irradiation

Published on: March 20, 2018

9.8K

SET8 localization to chromatin flanking DNA damage is dependent on RNF168 ubiquitin ligase

Stanimir Dulev1, Sichun Lin1, Qingquan Liu1

  • 1Department of Medical Biophysics, University of Toronto, Toronto, Canada.

Cell Cycle (Georgetown, Tex.)
|November 26, 2019
PubMed

Insights

The DNA damage response involves recruiting repair proteins to double-strand breaks (DSBs). This study reveals RNF168 ubiquitin ligase mediates SET8 methyltransferase recruitment to DSBs, a crucial step for DNA repair.

Area of Science:

  • Molecular Biology
  • Cellular Biology
  • Genetics

Background:

  • The DNA damage response (DDR) orchestrates the recruitment of chromatin remodelers and repair factors to DNA double-strand breaks (DSBs).
  • Proteins like 53BP1 are critical for DSB repair, and their localization depends on ubiquitin ligases (e.g., RNF168) and methyltransferases (e.g., SET8).

Purpose of the Study:

  • To investigate the precise mechanism by which RNF168 and SET8 contribute to 53BP1 recruitment at DSBs.
  • To determine if RNF168 and SET8 utilize the same pathways for their localization to DSBs.

Main Methods:

  • Depletion of cellular ubiquitin via proteasome inhibition.
  • Knockdown of RNF8 and RNF168.
  • Assessment of RNF168 and SET8 localization to DNA damage sites.
  • Analysis of RNF168 and SET8 complex formation in vivo.

Main Results:

  • RNF168 mediates the recruitment of SET8 to DSBs.
  • Depletion of ubiquitin or knockdown of RNF8/RNF168 prevents SET8 localization to DNA damage.
  • RNF168 and SET8 form stable complexes in vivo, suggesting a direct interaction.

Conclusions:

  • RNF168 is essential for recruiting SET8 to DSBs, clarifying a key step in the DDR pathway.
  • A model is proposed where SET8 accumulates at DSBs via tethering to proteins that recognize specific chromatin modifications, facilitated by RNF168.
Keywords:
DNA damage responseDNA repairRNF168SET8

More Related Videos

Immunofluorescence Imaging of DNA Damage and Repair Foci in Human Colon Cancer Cells05:18

Immunofluorescence Imaging of DNA Damage and Repair Foci in Human Colon Cancer Cells

Published on: June 9, 2020

11.7K
Capturing Common Fragile Site Breaks by Native γH2A.X ChIP09:46

Capturing Common Fragile Site Breaks by Native γH2A.X ChIP

Published on: January 24, 2025

659

Related Concept Videos

Nucleosome Remodeling02:54

Nucleosome Remodeling

Nucleosomes are the basic units of chromatin compaction. Each nucleosome consists of the DNA bound tightly around a histone core, which makes the DNA inaccessible to DNA binding proteins such as DNA polymerase and RNA polymerase. Hence, the fundamental problem is to ensure access to DNA when appropriate, despite the compact and protective chromatin structure.
Nucleosome remodeling complex
Eukaryotic cells have specialized enzymes called ATP-dependent nucleosome remodeling enzymes. These enzymes...
10.7K
Nucleotide Excision Repair01:38

Nucleotide Excision Repair

DNA Distortion and Damage
Cells are regularly exposed to mutagens—factors in the environment that can damage DNA and generate mutations. UV radiation is one of the most common mutagens and is estimated to introduce a significant number of changes in DNA. These include bends or kinks in the structure, which can block DNA replication or transcription. If these errors are not fixed, the damage can cause mutations, which in turn can result in cancer or disease depending on which sequences are...
4.9K
Nucleotide Excision Repair01:08

Nucleotide Excision Repair

Overview
40.5K
Homologous Recombination02:31

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
62.2K
DNA Damage can Stall the Cell Cycle02:36

DNA Damage can Stall the Cell Cycle

In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
9.9K
DNA Damage Can Stall the Cell Cycle02:36

DNA Damage Can Stall the Cell Cycle

In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
3.0K