Related Experiment Video
Updated: Apr 5, 2026
13:38
Monitoring Kinase and Phosphatase Activities Through the Cell Cycle by Ratiometric FRET
Published on: January 27, 2012
14.8K
Spatiotemporal Investigation of Phosphorylation Events During Cell Cycle Progression
Lilia Gheghiani1, Olivier Gavet
1Sorbonne Universités, UPMC Paris VI, UFR927, 75005, Paris, France.
Methods in Molecular Biology (Clifton, N.J.)
|August 10, 2015
Summary
Polo-like kinase 1 (Plk1) regulates cell cycle progression. Researchers developed a biosensor to visualize Cdc25C phosphorylation, revealing Plk1
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Polo-like kinase 1 (Plk1) is crucial for cell division.
- Plk1's roles in G2/M phase and Cdc25C regulation are not fully understood.
- Cdc25C phosphatase activates Cyclin B1-Cdk1, the main driver of mitosis.
Purpose of the Study:
- To investigate Plk1-dependent regulation of Cdc25C during G2/M progression.
- To develop and utilize a biosensor for real-time monitoring of Cdc25C phosphorylation.
- To provide a framework for creating biosensors for other phosphorylation sites.
Main Methods:
- Genetically encoded Förster Resonance Energy Transfer (FRET)-based biosensor development.
- Live single-cell assays to observe spatiotemporal phosphorylation.
- Cell cycle analysis.
Main Results:
- The FRET biosensor successfully visualized spatiotemporal Cdc25C phosphorylation.
- The study provided insights into Plk1's regulatory mechanisms during G2/M transition.
- The developed biosensor approach is adaptable for studying other phosphorylation events.
Conclusions:
- Plk1 plays a key role in regulating Cdc25C phosphorylation during the G2/M transition.
- The FRET biosensor is a powerful tool for studying cell cycle dynamics.
- Guidelines are provided for developing novel phosphorylation biosensors.
Related Concept Videos
02:54Protein Kinases and Phosphatases
15.6K
Proteins undergo chemical modifications that trigger changes in the charge, structure, and conformation of the proteins. Phosphorylation, acetylation, glycosylation, nitrosylation, ubiquitination, lipidation, methylation, and proteolysis are various protein modifications that regulate protein activity. Such modifications are usually enzyme-driven.
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
15.6K
02:39Positive Regulator Molecules
7.1K
Mitotic cell division results in daughter cells that exactly resemble the parent cell. However, errors in the DNA replication or distribution of genetic material may lead to genetic mutations that may be passed down to every new cell formed from the resulting abnormal cell. Propagation of such mutant cells is restricted through checkpoint mechanisms present at different stages of the cell cycle. These checkpoints involve regulator molecules that either promote or demote cell cycle events.
7.1K
01:45Positive Regulator Molecules
137.2K
To consistently produce healthy cells, the cell cycle—the process that generates daughter cells—must be precisely regulated.
137.2K
01:02Phosphorylation
55.7K
The addition or removal of phosphate groups from proteins is the most common chemical modification that regulates cellular processes. These modifications can affect the structure, activity, stability, and localization of proteins within cells as well as their interactions with other proteins.
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...
55.7K
02:15M-Cdk Drives Transition Into Mitosis
6.8K
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...
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...
6.8K
02:34Inhibition of Cdk Activity
6.2K
The orderly progression of the cell cycle depends on the activation of Cdk protein by binding to its cyclin partner. However, the cell cycle must be restricted when undergoing abnormal changes. Most cancers correlate to the deregulated cell cycle, and since Cdks are a central component of the cell cycle, Cdk inhibitors are extensively studied to develop anticancer agents. For instance, cyclin D associates with several Cdks, such as Cdk 4/6, to form an active complex. The cyclin D-Cdk4/6 complex...
6.2K