Related Experiment Video

Updated: Dec 31, 2025

Genetic Studies of Human DNA Repair Proteins Using Yeast as a Model System
14:09

Genetic Studies of Human DNA Repair Proteins Using Yeast as a Model System

Published on: March 18, 2010

18.4K

The Aspartic Protease Ddi1 Contributes to DNA-Protein Crosslink Repair in Yeast

Nataliia Serbyn1, Audrey Noireterre1, Ivona Bagdiul1

  • 1Department of Cell Biology, University of Geneva, 1211 Geneva 4, Switzerland.

Molecular Cell
|January 7, 2020
PubMed

Insights

DNA-protein crosslinks (DPCs) pose a threat to DNA transactions. The protease Ddi1 aids in repairing these lesions by degrading crosslinked proteins, working alongside Wss1/SPRTN.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • DNA-protein crosslinks (DPCs) are toxic lesions that impede DNA replication and transcription.
  • The protease Wss1/SPRTN is known to degrade DPCs, including topoisomerase-1 cleavage complexes (Top1ccs).
  • The precise mechanisms and additional factors involved in DPC repair are not fully understood.

Purpose of the Study:

  • To identify novel factors involved in DNA-protein crosslink (DPC) repair.
  • To elucidate the role of the predicted protease Ddi1 in Saccharomyces cerevisiae DPC processing.
  • To investigate the relationship between Ddi1, Wss1, and Top1cc repair.

Main Methods:

  • Genetic screening of yeast mutants defective in Top1cc processing.
  • Analysis of Ddi1 recruitment to DPC lesions.
  • Assessment of cell sensitivity to DPC-trapping agents.
  • Investigation of Ddi1's effect on protein degradation.

Main Results:

  • Ddi1 was identified in a genetic screen for mutants with defects in Top1cc processing.
  • Ddi1 is recruited to DPC lesions in an S phase-dependent manner to facilitate protein eviction.
  • Loss of Ddi1 function leads to hypersensitivity to DPC agents, independent of Wss1 and the proteasome.
  • Ddi1's protease activity is crucial for the degradation of crosslinked proteins, including a core component of RNA Polymerase II.

Conclusions:

  • Ddi1 is a crucial protease involved in the efficient disassembly of DNA-protein crosslinks (DPCs).
  • Ddi1 acts in concert with Wss1/SPRTN to repair DPCs, expanding the known repertoire of DPC proteases.
  • Ddi1 plays a significant role in maintaining genome stability by promoting the repair of genotoxin-induced DNA damage.
Keywords:
26S proteasomeDNA-protein crosslinkDdi1Flp recombinaseRNA polymerase IIRpb1Top1Wss1proteaseyeast

More Related Videos

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae07:55

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae

Published on: September 11, 2022

2.1K
Author Spotlight: Quantitative Detection of DNA Protein Crosslinks and Their Post-Translational Modifications10:12

Author Spotlight: Quantitative Detection of DNA Protein Crosslinks and Their Post-Translational Modifications

Published on: April 21, 2023

3.4K

Related Concept Videos

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...
62.1K
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
Fixing Double-strand Breaks02:04

Fixing Double-strand Breaks

The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...
14.2K
Base-pairing and DNA Repair02:27

Base-pairing and DNA Repair

89.6K
Protein Modifications in the RER01:26

Protein Modifications in the RER

Modification of secretory and transmembrane proteins entering the rough ER begins in the ER lumen. These modifications aid in protein folding and stabilize the acquired tertiary structure. Protein modifications in the rough ER co-occur at different stages of protein folding.
Broadly, these modifications can be categorized into four main categories — glycosylation, formation of disulfide bonds, assembly of protein subunits, and specific proteolytic cleavages like removal of signal...
6.7K