Related Experiment Video

Updated: Aug 9, 2026

Two- and Three-Dimensional Live Cell Imaging of DNA Damage Response Proteins
10:24

Two- and Three-Dimensional Live Cell Imaging of DNA Damage Response Proteins

Published on: September 28, 2012

Mec1p is essential for phosphorylation of the yeast DNA damage checkpoint protein Ddc1p, which physically interacts

V Paciotti1, G Lucchini, P Plevani

  • 1Dipartimento di Genetica e di Biologia dei Microrganismi, Università degli Studi di Milano, Via Celoria 26, 20133 Milano, Italy.

The EMBO Journal
|July 22, 1998
PubMed

Insights

The Saccharomyces cerevisiae DDC1 gene is crucial for DNA damage checkpoints. Ddc1p interacts with Mec3p and is phosphorylated by Mec1p, suggesting a role in early DNA damage response pathways.

Area of Science:

  • Cellular biology
  • Molecular genetics
  • DNA damage response

Background:

  • DNA checkpoints prevent cell division with damaged chromosomes.
  • The Saccharomyces cerevisiae DDC1 gene is part of the RAD17, MEC3, and RAD24 epistasis group, proposed to function early in DNA damage checkpoint pathways.
  • Ddc1p exhibits cell cycle-dependent phosphorylation, increasing upon DNA damage.

Purpose of the Study:

  • To investigate the physical interactions and phosphorylation dependencies of Ddc1p.
  • To elucidate the role of Ddc1p within the DNA damage checkpoint pathway.

Main Methods:

  • In vivo physical interaction studies (co-immunoprecipitation).
  • Analysis of Ddc1p phosphorylation in various mutant backgrounds.
  • Investigating Ddc1p's role in the phosphorylation of other checkpoint proteins.

Main Results:

  • Ddc1p physically interacts with Mec3p in vivo, requiring Rad17p.
  • Ddc1p phosphorylation is dependent on Mec1p, Rad24p, Rad17p, and Mec3p, but independent of Rad53p and Rad9p.
  • Ddc1p is necessary for Rad53p phosphorylation but not for Pds1p phosphorylation.

Conclusions:

  • Mec1p and the Rad24 protein group likely collaborate in early DNA damage checkpoint signaling.
  • Ddc1p and Rad9p may operate in parallel branches of the DNA damage response, influencing Mec1p activity and substrate specificity.

More Related Videos

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay12:26

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay

Published on: May 3, 2018

Use of Time-Lapse Microscopy and Stage-Specific Nuclear Depletion of Proteins to Study Meiosis in S. cerevisiae07:48

Use of Time-Lapse Microscopy and Stage-Specific Nuclear Depletion of Proteins to Study Meiosis in S. cerevisiae

Published on: October 11, 2022

Related Concept Videos

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...
M-Cdk Drives Transition Into Mitosis02:15

M-Cdk Drives Transition Into Mitosis

Checkpoints throughout the cell cycle serve as safeguards and gatekeepers, allowing the cell cycle to progress in favorable conditions and slow or halt it in problematic ones. This regulation is known as the cell cycle control system.
Cyclin-dependent kinases, or Cdks, work in concert with cyclins to control cell cycle transitions. M-Cdk, a complex of Cdk1 bound to M cyclin, is a well-known example of this coordinated control that drives the transition from the G2 to the M phase.
M cyclin...
Separation of Sister Chromatids02:17

Separation of Sister Chromatids

At the transition from prophase to metaphase, there is a reduction in cohesion along the chromosomal arms, resulting in the resolution of sister chromatids. However, residual cohesin connections remain to hold the sister chromatids together until the transition from metaphase to anaphase. The residual connection prevents any premature separation of sister chromatids, blocking the risks of aneuploidy within the daughter cells.
At the onset of anaphase, separase, a proteolytic enzyme, is...
The Spindle Assembly Checkpoint02:19

The Spindle Assembly Checkpoint

The spindle assembly checkpoint is a molecular surveillance mechanism ensuring the fidelity of chromosome segregation during anaphase. The checkpoint monitors the completion of all the prerequisite steps before chromosome segregation to determine whether the segregation process should proceed or be delayed.
Many proteins function together to control the spindle assembly checkpoint. Mutations affecting these proteins may allow cells to proceed into anaphase prematurely, resulting in the...
Anaphase Promoting Complex00:50

Anaphase Promoting Complex

The stepwise destruction of specific proteins is necessary for the progression and completion of the cell cycle. Such proteins are ubiquitinated by ubiquitin ligases and then subsequently destroyed by the proteasome. The SCF (Skp1/Cullin/F-box) and the anaphase-promoting complex (APC) are two important ubiquitin ligases involved in cell cycle progression. While SCF is active throughout the cell cycle, APC gets activated during metaphase to anaphase transition. Cdc20 or Cdh1 binds to APC and...
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...