Related Experiment Video
Updated: Sep 6, 2025
10:54
Studying Proteolysis of Cyclin B at the Single Cell Level in Whole Cell Populations
Published on: September 17, 2012
10.6K
Cell cycle-dependent binding between Cyclin B1 and Cdk1 revealed by time-resolved fluorescence correlation
Martina Barbiero1, Luca Cirillo1, Sapthaswaran Veerapathiran1
1Cancer Biology, The Institute of Cancer Research Chester Beatty Laboratories, 237 Fulham Road, London, London SW3 6JB, UK.
Open Biology
|June 29, 2022
Summary
Understanding cell cycle control requires measuring protein complex dynamics. New microscopy and gene editing methods allow studying protein interactions in living cells, revealing Cyclin B1 and Cdk1 complex regulation during the cell cycle.
Area of Science:
- Cell Biology
- Biochemistry
- Molecular Dynamics
Background:
- Cell cycle control relies on precise regulation of protein complex assembly and disassembly.
- Measuring these dynamics in living cells has been challenging due to limitations of traditional biochemical and single-cell techniques.
Purpose of the Study:
- To investigate the dynamic assembly and disassembly of cell cycle regulatory complexes in living cells.
- To quantify the kinetics of protein-protein interactions within the cell cycle machinery.
Main Methods:
- Application of fluorescence correlation spectroscopy (FCS) and fluorescence cross-correlation spectroscopy (FCCS).
- Utilizing advanced microscopy and gene editing techniques for real-time measurements in living cells.
Main Results:
- Identified a significant pool of unbound Cyclin B1 in living cells, despite known high-affinity binding to Cdk1.
- Demonstrated an increase in Cyclin B1-Cdk1 binding affinity throughout the cell cycle.
- Provided evidence that Cyclin B1-Cdk1 complex assembly is a regulated process.
Conclusions:
- The assembly of the Cyclin B1-Cdk1 complex, a key regulator of mitosis, is a dynamic and regulated step in the cell cycle.
- This study establishes a foundation for future investigations into the kinetics of protein complex dynamics during cell division.
More Related Videos
Related Concept Videos
02:39Positive Regulator Molecules
5.6K
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.
5.6K
02:34Inhibition of Cdk Activity
4.9K
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...
4.9K
01:19Protein Dynamics in Living Cells
2.2K
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
2.2K
02:15M-Cdk Drives Transition Into Mitosis
5.7K
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...
5.7K