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Related Concept Videos

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.
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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...
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...
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...

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Related Experiment Video

Updated: Jul 25, 2026

Studying Proteolysis of Cyclin B at the Single Cell Level in Whole Cell Populations
10:54

Studying Proteolysis of Cyclin B at the Single Cell Level in Whole Cell Populations

Published on: September 17, 2012

Separate SCF(CDC4) recognition elements target Cdc6 for proteolysis in S phase and mitosis.

G Perkins1, L S Drury, J F Diffley

  • 1ICRF Clare Hall Laboratories, South Mimms EN6 3LD, UK.

The EMBO Journal
|September 5, 2001
PubMed
Summary

A novel dominant-negative Cdc6 mutant arrests the cell cycle by inhibiting cyclin-dependent kinases (CDKs). This mutant is resistant to proteolysis, leading to cell cycle arrest and providing insights into DNA replication regulation.

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Combining Mitotic Cell Synchronization and High Resolution Confocal Microscopy to Study the Role of Multifunctional Cell Cycle Proteins During Mitosis
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Manipulation and Analysis of Cell Cycle-Dependent Processes in Budding Yeast
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Related Experiment Videos

Last Updated: Jul 25, 2026

Studying Proteolysis of Cyclin B at the Single Cell Level in Whole Cell Populations
10:54

Studying Proteolysis of Cyclin B at the Single Cell Level in Whole Cell Populations

Published on: September 17, 2012

Combining Mitotic Cell Synchronization and High Resolution Confocal Microscopy to Study the Role of Multifunctional Cell Cycle Proteins During Mitosis
08:33

Combining Mitotic Cell Synchronization and High Resolution Confocal Microscopy to Study the Role of Multifunctional Cell Cycle Proteins During Mitosis

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Manipulation and Analysis of Cell Cycle-Dependent Processes in Budding Yeast
08:13

Manipulation and Analysis of Cell Cycle-Dependent Processes in Budding Yeast

Published on: September 26, 2025

Area of Science:

  • Molecular Biology
  • Cell Cycle Regulation
  • Yeast Genetics

Background:

  • Cdc6 is a key DNA replication initiation factor in Saccharomyces cerevisiae.
  • Cdc6 is regulated by ubiquitin-mediated proteolysis via the SCF(CDC4) E3 ubiquitin ligase.
  • SCF(CDC4)-mediated proteolysis targets Cdc6 from late G1 phase through mitosis.

Purpose of the Study:

  • To characterize a dominant-negative CDC6 mutant.
  • To investigate the mechanism by which this mutant inhibits cell cycle progression.
  • To identify novel regulatory sites on Cdc6 involved in proteolysis and cell cycle control.

Main Methods:

  • Overexpression of a dominant-negative CDC6 mutant.
  • Cell cycle arrest analysis.
  • Analysis of cyclin-dependent kinase (CDK) inhibition.
  • Investigation of SCF(CDC4)-mediated proteolysis.
  • Site-directed mutagenesis to identify phosphorylation sites and interaction domains.

Main Results:

  • The dominant-negative CDC6 mutant arrests the cell cycle by inhibiting CDKs.
  • The mutant protein is refractory to SCF(CDC4)-mediated proteolysis, leading to its accumulation.
  • A mutation destroying a putative CDK phosphorylation site confers the dominant-negative phenotype.
  • This mutation affects a novel Cdc4-interacting domain, distinct from a previously identified N-terminal site.
  • Both identified sites regulate Cdc6 proteolysis, but the novel site is crucial during mitosis.

Conclusions:

  • A novel dominant-negative CDC6 mutant inhibits cell cycle progression by interfering with CDK activity.
  • The resistance of this mutant to proteolysis is key to its accumulation and cell cycle arrest.
  • A newly identified Cdc4-interacting domain and CDK phosphorylation site are critical for Cdc6 regulation, particularly during mitosis.