Tension-dependent regulation of microtubule dynamics at kinetochores can explain metaphase congression in yeast

During metaphase in budding yeast mitosis, sister kinetochores are tethered to opposite poles and separated, stretching their intervening chromatin, by singly attached kinetochore microtubules (kMTs). Kinetochore movements are coupled to single microtubule plus-end polymerization/depolymerization at...

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Bibliographic Details
Published in:Molecular biology of the cell 2005-08, Vol.16 (8), p.3764-3775
Main Authors: Gardner, Melissa K, Pearson, Chad G, Sprague, Brian L, Zarzar, Ted R, Bloom, Kerry, Salmon, E D, Odde, David J
Format: Article
Language:English
Subjects:
Cell Cycle Proteins - genetics
Cell Cycle Proteins - metabolism
Chromatin - metabolism
Chromosomal Proteins, Non-Histone
DNA-Binding Proteins - metabolism
Kinetochores - metabolism
Metaphase
Microtubules - metabolism
Models, Biological
Mutation - genetics
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
Saccharomyces cerevisiae
Saccharomyces cerevisiae - cytology
Saccharomyces cerevisiae - metabolism
Saccharomyces cerevisiae Proteins - genetics
Saccharomyces cerevisiae Proteins - metabolism
Spectrometry, Fluorescence
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Summary:During metaphase in budding yeast mitosis, sister kinetochores are tethered to opposite poles and separated, stretching their intervening chromatin, by singly attached kinetochore microtubules (kMTs). Kinetochore movements are coupled to single microtubule plus-end polymerization/depolymerization at kinetochore attachment sites. Here, we use computer modeling to test possible mechanisms controlling chromosome alignment during yeast metaphase by simulating experiments that determine the 1) mean positions of kinetochore Cse4-GFP, 2) extent of oscillation of kinetochores during metaphase as measured by fluorescence recovery after photobleaching (FRAP) of kinetochore Cse4-GFP, 3) dynamics of kMTs as measured by FRAP of GFP-tubulin, and 4) mean positions of unreplicated chromosome kinetochores that lack pulling forces from a sister kinetochore. We rule out a number of possible models and find the best fit between theory and experiment when it is assumed that kinetochores sense both a spatial gradient that suppresses kMT catastrophe near the poles and attachment site tension that promotes kMT rescue at higher amounts of chromatin stretch.
ISSN:1059-1524
1939-4586
1059-1524
DOI:10.1091/mbc.E05-04-0275