Measuring nanometer scale gradients in spindle microtubule dynamics using model convolution microscopy

Chad G. Pearson, Melissa K Gardner, Leocadia V. Paliulis, E. D. Salmon, David J Odde, Kerry Bloom

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

A computational model for the budding yeast mitotic spindle predicts a spatial gradient in tubulin turnover that is produced by kinetochore-attached microtubule (kMT) plus-end polymerization and depolymerization dynamics. However, kMTs in yeast are often much shorter than the resolution limit of the light microscope, making visualization of this gradient difficult. To overcome this limitation, we combined digital imaging of fluorescence redistribution after photobleaching (FRAP) with model convolution methods to compare computer simulations at nanometer scale resolution to microscopic data. We measured a gradient in microtubule dynamics in yeast spindles at ∼65-nm spatial intervals. Tubulin turnover is greatest near kinetochores and lowest near the spindle poles. A β-tubulin mutant with decreased plus-end dynamics preserves the spatial gradient in tubulin turnover at a slower time scale, increases average kinetochore microtubule length ∼14%, and decreases tension at kinetochores. The β-tubulin mutant cells have an increased frequency of chromosome loss, suggesting that the accuracy of chromosome segregation is linked to robust kMT plus-end dynamics.

Original languageEnglish (US)
Pages (from-to)4069-4079
Number of pages11
JournalMolecular Biology of the Cell
Volume17
Issue number9
DOIs
StatePublished - Sep 1 2006

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Kinetochores
Tubulin
Microtubules
Microscopy
Yeasts
Spindle Poles
Photobleaching
Chromosome Segregation
Spindle Apparatus
Saccharomycetales
Optical Imaging
Polymerization
Computer Simulation
Chromosomes
Light

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Measuring nanometer scale gradients in spindle microtubule dynamics using model convolution microscopy. / Pearson, Chad G.; Gardner, Melissa K; Paliulis, Leocadia V.; Salmon, E. D.; Odde, David J; Bloom, Kerry.

In: Molecular Biology of the Cell, Vol. 17, No. 9, 01.09.2006, p. 4069-4079.

Research output: Contribution to journalArticle

Pearson, Chad G. ; Gardner, Melissa K ; Paliulis, Leocadia V. ; Salmon, E. D. ; Odde, David J ; Bloom, Kerry. / Measuring nanometer scale gradients in spindle microtubule dynamics using model convolution microscopy. In: Molecular Biology of the Cell. 2006 ; Vol. 17, No. 9. pp. 4069-4079.
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