Related Experiment Video

Updated: Feb 1, 2026

Chromatin Interaction Analysis with Paired-End Tag Sequencing ChIA-PET for Mapping Chromatin Interactions and Understanding Transcription Regulation
21:55

Chromatin Interaction Analysis with Paired-End Tag Sequencing ChIA-PET for Mapping Chromatin Interactions and Understanding Transcription Regulation

Published on: April 30, 2012

31.2K

The OTUD5-UBR5 complex regulates FACT-mediated transcription at damaged chromatin

Angelo de Vivo1, Anthony Sanchez1, Jose Yegres1

  • 1Department of Cell Biology, Microbiology, and Molecular Biology, College of Arts and Sciences, University of South Florida, Tampa, FL 33620, USA.

Nucleic Acids Research
|December 4, 2018
PubMed

Insights

OTUD5 deubiquitinase stabilizes UBR5 and binds FACT, halting RNA Pol II at DNA breaks. This reveals OTUD5

Area of Science:

  • Molecular Biology
  • Chromatin Biology
  • DNA Damage Response

Background:

  • RNA polymerase (Pol II) stalling and resumption at damaged chromatin are crucial regulated processes.
  • The FACT histone chaperone complex is known to play a role in managing chromatin during transcription.
  • The deubiquitinase OTUD5's function in these processes was previously unknown.

Purpose of the Study:

  • To investigate the role of OTUD5 in the FACT-dependent regulation of transcription at damaged chromatin.
  • To identify new regulators of DNA double-strand break (DSB) response pathways.

Main Methods:

  • RNA interference (RNAi) screen for deubiquitinating enzymes (DUBs).
  • Immunoprecipitation and co-localization studies to analyze protein interactions.
  • Analysis of RNA Pol II activity and RNA synthesis at DSB sites.
  • Site-directed mutagenesis to study the functional impact of specific mutations.

Main Results:

  • OTUD5 was identified as a specific stabilizer of the UBR5 E3 ligase.
  • OTUD5 localizes to DSBs, interacts with UBR5 and SPT16 (a FACT component), and represses RNA Pol II elongation.
  • OTUD5 antagonizes H2A deposition at DSB lesions and its interactions with UBR5 and SPT16 are essential for Pol II arrest.
  • A cancer-associated mutation in OTUD5's UIM disrupts FACT association and Pol II arrest.

Conclusions:

  • OTUD5 acts as a novel regulator in the DNA damage response by stabilizing UBR5 and interacting with FACT.
  • Both catalytic (via UBR5) and scaffolding (via FACT) functions of OTUD5 are critical for FACT-dependent transcription regulation at damaged sites.
  • Dysregulation of OTUD5, potentially through mutations, may link transcriptional control to tumor suppression.

More Related Videos

Sequential Salt Extractions for the Analysis of Bulk Chromatin Binding Properties of Chromatin Modifying Complexes07:41

Sequential Salt Extractions for the Analysis of Bulk Chromatin Binding Properties of Chromatin Modifying Complexes

Published on: October 2, 2017

8.9K
Describing a Transcription Factor Dependent Regulation of the MicroRNA Transcriptome07:23

Describing a Transcription Factor Dependent Regulation of the MicroRNA Transcriptome

Published on: June 15, 2016

8.9K

Related Concept Videos

Chromatin Structure Regulates pre-mRNA Processing02:41

Chromatin Structure Regulates pre-mRNA Processing

In eukaryotic cells, nascent mRNA transcripts need to undergo many post-transcriptional modifications to reach the cell cytoplasm and translate into functional proteins. For a long time, transcription and pre-mRNA processing were considered two independent events that occur sequentially in the cell. However, it has now been well established that transcription and pre-mRNA processing are two simultaneous processes that are precisely regulated inside the cell.
The chromatin structure, especially...
8.2K
Master Transcription Regulators02:23

Master Transcription Regulators

Master transcription regulators are regulatory proteins that are predominantly responsible for regulating the expression of multiple genes. Often these genes work in concert to drive a  complex process. Activation of a master transcription regulator can lead to a cascade of transcriptional activation necessary for that outcome. These regulators can directly bind to the regulatory sequences of the various genes involved, or they can indirectly regulate transcription by binding to regulatory...
7.8K
Master Transcription Regulators02:23

Master Transcription Regulators

2.8K
Cooperative Binding of Transcription Regulators02:13

Cooperative Binding of Transcription Regulators

Transcriptional regulators bind to specific cis-regulatory sequences in the DNA to regulate gene transcription. These cis-regulatory sequences are very short, usually less than ten nucleotide pairs in length. The short length means that there is a high probability of the exact same sequence randomly occurring throughout the genome.  Since regulators can also bind to groups of similar sequences, this further increases the chances of random binding. Transcriptional regulators form...
7.3K
Spreading of Chromatin Modifications02:25

Spreading of Chromatin Modifications

The histone proteins in the nucleosomes are post-translationally modified (PTM) to increase or decrease access to DNA. The commonly observed PTMs are methylation, acetylation, phosphorylation, and ubiquitination of lysine amino acids in the histone H3 tail region. These histone modifications have specific meaning for the cell. Hence, they are called "histone code". The protein complex involved in histone modification is termed as "reader-writer" complex.
Writers
The writer...
9.5K
Transcription Factors02:16

Transcription Factors

Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
82.7K