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BiFCo: visualizing cohesin assembly/disassembly cycle in living cells
Emilio González-Martín1, Juan Jiménez2, Víctor A Tallada3
1Centro Andaluz de Biología del Desarrollo, Universidad Pablo de Olavide-Consejo Superior de Investigaciones Científicas-Junta de Andalucía, Seville, Spain.
Life Science Alliance
|May 9, 2023
Summary
Researchers developed bimolecular fluorescent cohesin (BiFCo) to visualize cohesin complex dynamics in living cells. This method overcomes challenges posed by excess soluble proteins, enabling study of cohesin assembly and disassembly.
Area of Science:
- Cell Biology
- Molecular Biology
- Genetics
Background:
- Cohesin, a ring-shaped complex of SMC proteins and RAD21, is crucial for genome stability in eukaryotes.
- Mutations in cohesin lead to cohesinopathies and cancer.
- Investigating cohesin dynamics in vivo is challenging due to excess soluble subunits, hindering fluorescent fusion studies.
Purpose of the Study:
- To develop a novel method for visualizing cohesin complex assembly and disassembly in living cells.
- To overcome the limitations of traditional fluorescent fusion techniques for studying cohesin dynamics.
Main Methods:
- Utilized bimolecular fluorescent complementation (BiFCo) in the fission yeast Schizosaccharomyces pombe.
- Designed BiFCo to selectively detect assembled cohesin complexes, excluding signals from individual subunits.
Main Results:
- BiFCo enables monitoring of cohesin complex assembly and disassembly within a physiological context.
- The system allows observation throughout the entire cell cycle in living cells.
- Demonstrated the ability to track dynamic changes in cohesin complex formation.
Conclusions:
- BiFCo provides a versatile tool for studying cohesin dynamics in real-time.
- This system can be adapted for various genetic backgrounds and other eukaryotic models, including human cells.
- Offers new avenues for understanding cohesinopathies and related diseases.
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