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Biomolecules 2019, 9(3), 86; https://doi.org/10.3390/biom9030086

Quantifying Tubulin Concentration and Microtubule Number Throughout the Fission Yeast Cell Cycle

1
Cell & Developmental Biology, University of Pennsylvania, Philadelphia, PA 19104, USA
2
Molecular Devices Corporation, Downingtown, PA 19335, USA
3
Institut Curie, PSL Research University, CNRS, UMR 144, F-75005 Paris, France
*
Author to whom correspondence should be addressed.
Received: 19 December 2018 / Revised: 25 February 2019 / Accepted: 26 February 2019 / Published: 4 March 2019
(This article belongs to the Special Issue Cytoskeleton and Regulation of Mitosis)
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Abstract

The fission yeast Schizosaccharomyces pombe serves as a good genetic model organism for the molecular dissection of the microtubule (MT) cytoskeleton. However, analysis of the number and distribution of individual MTs throughout the cell cycle, particularly during mitosis, in living cells is still lacking, making quantitative modelling imprecise. We use quantitative fluorescent imaging and analysis to measure the changes in tubulin concentration and MT number and distribution throughout the cell cycle at a single MT resolution in living cells. In the wild-type cell, both mother and daughter spindle pole body (SPB) nucleate a maximum of 23 ± 6 MTs at the onset of mitosis, which decreases to a minimum of 4 ± 1 MTs at spindle break down. Interphase MT bundles, astral MT bundles, and the post anaphase array (PAA) microtubules are composed primarily of 1 ± 1 individual MT along their lengths. We measure the cellular concentration of αβ-tubulin subunits to be ~5 µM throughout the cell cycle, of which one-third is in polymer form during interphase and one-quarter is in polymer form during mitosis. This analysis provides a definitive characterization of αβ-tubulin concentration and MT number and distribution in fission yeast and establishes a foundation for future quantitative comparison of mutants defective in MTs. View Full-Text
Keywords: microtubule dynamics; spindle morphogenesis microtubule dynamics; spindle morphogenesis
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Loiodice, I.; Janson, M.E.; Tavormina, P.; Schaub, S.; Bhatt, D.; Cochran, R.; Czupryna, J.; Fu, C.; Tran, P.T. Quantifying Tubulin Concentration and Microtubule Number Throughout the Fission Yeast Cell Cycle. Biomolecules 2019, 9, 86.

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