Related Experiment Video
Updated: Apr 19, 2026
07:55
Visualization of DNA Repair Proteins Interaction by Immunofluorescence
Published on: June 26, 2020
11.4K
BRCA2 diffuses as oligomeric clusters with RAD51 and changes mobility after DNA damage in live cells
Marcel Reuter1, Alex Zelensky1, Ihor Smal2
1Department of Genetics, Cancer Genomics Centre Netherlands, Department of Medical Informatics, Department of Radiology, Erasmus Optical Imaging Centre, Department of Pathology, Department of Vascular Surgery, and Department of Radiation Oncology, Erasmus University Medical Centre, 3000 CA Rotterdam, Netherlands.
The Journal of Cell Biology
|December 10, 2014
Summary
Researchers tracked BRCA2 protein mobility in live cells to understand its role in DNA repair. They found BRCA2 interacts with RAD51, suggesting BRCA2 sequesters RAD51 to prevent DNA damage and deliver it to break sites.
Area of Science:
- Molecular Biology
- Genetics
- Cell Biology
Background:
- Genome maintenance relies on homologous recombination, a process involving coordinated protein assembly at DNA break sites.
- BRCA2 is a critical mediator protein in homologous recombination, essential for DNA repair fidelity.
Purpose of the Study:
- To investigate the dynamic behavior and mobility of BRCA2 in live cells at the single-molecule level.
- To elucidate the interaction between BRCA2 and RAD51 during DNA repair processes.
Main Methods:
- Single-particle tracking and fluorescence correlation spectroscopy were employed to monitor fluorescently tagged BRCA2, RAD51, and RAD54 proteins.
- Proteins were expressed from endogenous loci to reflect physiological conditions, with comparisons made between BRCA2-GFP and -YFP to account for fluorophore effects.
Main Results:
- Nuclear BRCA2 was observed to exist in oligomeric clusters with heterogeneous mobility, which increased transient binding upon DNA damage.
- RAD51 exhibited similar mobility to BRCA2, indicating a physical interaction that persists both before and after DNA damage induction.
- BRCA2-GFP and -YFP were used to differentiate protein diffusion from fluorophore behavior.
Conclusions:
- BRCA2 plays a crucial role in regulating RAD51 dynamics within the nucleus.
- BRCA2-mediated sequestration of RAD51 is proposed to prevent aberrant DNA interactions and ensure efficient delivery of RAD51 to DNA damage sites.
Related Concept Videos
02:04Fixing Double-strand Breaks
16.5K
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...
16.5K
02:36DNA Damage can Stall the Cell Cycle
10.4K
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...
10.4K
02:36DNA Damage Can Stall the Cell Cycle
3.4K
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...
3.4K
02:31Homologous Recombination
66.1K
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...
66.1K
01:38Nucleotide Excision Repair
6.0K
DNA Distortion and Damage
Cells are regularly exposed to mutagens—factors in the environment that can damage DNA and generate mutations. UV radiation is one of the most common mutagens and is estimated to introduce a significant number of changes in DNA. These include bends or kinks in the structure, which can block DNA replication or transcription. If these errors are not fixed, the damage can cause mutations, which in turn can result in cancer or disease depending on which sequences are...
Cells are regularly exposed to mutagens—factors in the environment that can damage DNA and generate mutations. UV radiation is one of the most common mutagens and is estimated to introduce a significant number of changes in DNA. These include bends or kinks in the structure, which can block DNA replication or transcription. If these errors are not fixed, the damage can cause mutations, which in turn can result in cancer or disease depending on which sequences are...
6.0K
01:18Other Unique Bacteria
569
Magnetic bacteria exhibit a directed movement called magnetotaxis, driven by structures called magnetosomes. These magnetosomes consist of chains of magnetic particles made of either magnetite (Fe₃O₄) or greigite (Fe₃S₄) and are organized in a linear conformation by a protein scaffold within invaginations of the cell membrane. The bacteria align along the north–south magnetic field lines, much like a compass needle. They are typically microaerophilic or anaerobic...
569