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Yeast kinetochore microtubule dynamics analyzed by high-resolution three-dimensional microscopy
J F Dorn1, K Jaqaman, D R Rines
1Laboratory for Computational Cell Biology, Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
Biophysical Journal
|September 30, 2005
Summary
Budding yeast chromosomes dynamically attach to microtubules in G1, unlike in metazoans. This study uses high-resolution microscopy to reveal kinetochore microtubule (k-MT) dynamics, offering a sensitive assay for k-MT regulation.
Area of Science:
- Cell Biology
- Microscopy
- Molecular Biology
Background:
- Kinetochore-microtubule (k-MT) attachments are crucial for chromosome segregation.
- Understanding k-MT dynamics in G1 is essential for cell cycle progression.
Purpose of the Study:
- To probe single k-MT dynamics in budding yeast G1 phase.
- To establish a sensitive assay for k-MT regulation using high-resolution light microscopy.
Main Methods:
- Automated tracking of fluorescent protein tags near centromeres and on spindle pole body proteins.
- Analysis of k-MT dynamics under various conditions (temperature, chemical treatments, mutations).
Main Results:
- Budding yeast chromosomes exhibit dynamic k-MT attachments in G1, differing from metazoan behavior.
- Lowering temperature and benomyl exposure similarly attenuate k-MT dynamics.
- The tub2-150 tubulin mutation leads to faster k-MT depolymerization than in wild-type cells.
Conclusions:
- Centromere dynamics in G1 yeast cells can serve as a sensitive indicator of k-MT regulation.
- The findings provide new insights into chromosome-microtubule interactions during the cell cycle.