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Updated: Jan 15, 2026
08:13
Manipulation and Analysis of Cell Cycle-Dependent Processes in Budding Yeast
Published on: September 26, 2025
473
Manipulation and Analysis of Cell Cycle-Dependent Processes in Budding Yeast.
Michael G Stewart1, Talia C Scheel2, Ahmed A Abouelghar2
1Department of Biochemistry, University of Utah School of Medicine; Department of Molecular Biology and Genetics, Howard Hughes Medical Institute, Johns Hopkins University School of Medicine; mstewa64@jh.edu.
Journal of Visualized Experiments : Jove
|October 13, 2025
Summary
This study presents robust methods for synchronizing budding yeast cell cycles, enabling detailed analysis of cell cycle-dependent processes. These techniques facilitate precise examination of protein dynamics and cellular events.
Area of Science:
- Cell Biology
- Molecular Biology
- Genetics
Background:
- Eukaryotic cell cycle regulation is crucial for DNA maintenance and organelle homeostasis.
- Cell cycle-dependent studies are vital for accurate interpretation of experimental data.
- Budding yeast is a powerful model organism for cell cycle research due to its genetic tractability and synchronization methods.
Purpose of the Study:
- To detail robust protocols for cell cycle synchronization in budding yeast.
- To enable the study of cell cycle-dependent protein localization and abundance changes.
- To provide methods adaptable for diverse cell cycle analyses, including live imaging and biochemical assays.
Main Methods:
- Synchronization of yeast cell cycles using G1 arrest with α-factor and mitotic arrest via Cdc20 depletion.
- Strain construction using PCR tagging for fluorescence microscopy and cell cycle arrests.
- Microscopy-based analysis of fixed cells, focusing on dynamic protein localization and population changes.
Main Results:
- Established protocols for G1 and mitotic arrest-release experiments in budding yeast.
- Demonstrated PCR tagging methods for creating strains suitable for cell cycle analysis.
- Enabled detailed imaging and analysis of protein dynamics throughout the cell cycle.
Conclusions:
- The presented synchronization methods are effective for studying cell cycle-dependent processes in yeast.
- These protocols can be adapted for various analyses, including live cell imaging and biochemical assays.
- Budding yeast remains an excellent model for advancing cell cycle research.
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