Mechanisms of microtubule-based kinetochore positioning in the yeast metaphase spindle.

@article{Sprague2003, abstract = {It has been hypothesized that spatial gradients in kMT dynamic instability facilitate mitotic spindle formation and chromosome movement. To test this hypothesis requires the analysis of kMT dynamics, which have not been resolved at the single kMT level in living cells. The budding yeast spindle offers an attractive system in which to study kMT dynamics because, in contrast to animal cells, there is only one kMT per kinetochore. To visualize metaphase kMT plus-end dynamics in yeast, a strain containing a green fluorescent protein fusion to the kinetochore protein, Cse4, was imaged by fluorescence microscopy. Although individual kinetochores were not resolvable, we found that models of kMT dynamics could be evaluated by simulating the stochastic kMT dynamics and then simulating the fluorescence imaging of kMT plus-end-associated kinetochores. Statistical comparison of model-predicted images to experimentally observed images demonstrated that a pure dynamic instability model for kMT dynamics in the yeast metaphase spindle was unacceptable. However, when a temporally stable spatial gradient in the catastrophe or rescue frequency was added to the model, there was reasonable agreement between the model and the experiment. These results provide the first evidence of temporally stable spatial gradients of kMT catastrophe and/or rescue frequency in living cells.}, address = {Department of Biomedical Engineering, University of Minnesota, Minneapolis, Minnesota 55455, USA.}, author = {Sprague, B. L. and Pearson, C. G. and Maddox, P. S. and Bloom, K. S. and Salmon, E. D. and Odde, D. J. }, citeulike-article-id = {2239611}, issn = {0006-3495}, journal = {Biophys J}, keywords = {kinetochore, positioning, yeast}, month = {June}, number = {6}, pages = {3529--3546}, posted-at = {2008-04-23 16:42:59}, priority = {0}, title = {Mechanisms of microtubule-based kinetochore positioning in the yeast metaphase spindle.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/12770865}, volume = {84}, year = {2003} }

TY - JOUR ID - Sprague2003 L3 - citeulike-article-id:2239611 N2 - It has been hypothesized that spatial gradients in kMT dynamic instability facilitate mitotic spindle formation and chromosome movement. To test this hypothesis requires the analysis of kMT dynamics, which have not been resolved at the single kMT level in living cells. The budding yeast spindle offers an attractive system in which to study kMT dynamics because, in contrast to animal cells, there is only one kMT per kinetochore. To visualize metaphase kMT plus-end dynamics in yeast, a strain containing a green fluorescent protein fusion to the kinetochore protein, Cse4, was imaged by fluorescence microscopy. Although individual kinetochores were not resolvable, we found that models of kMT dynamics could be evaluated by simulating the stochastic kMT dynamics and then simulating the fluorescence imaging of kMT plus-end-associated kinetochores. Statistical comparison of model-predicted images to experimentally observed images demonstrated that a pure dynamic instability model for kMT dynamics in the yeast metaphase spindle was unacceptable. However, when a temporally stable spatial gradient in the catastrophe or rescue frequency was added to the model, there was reasonable agreement between the model and the experiment. These results provide the first evidence of temporally stable spatial gradients of kMT catastrophe and/or rescue frequency in living cells. IS - 6 JF - Biophys J SN - 0006-3495 AD - Department of Biomedical Engineering, University of Minnesota, Minneapolis, Minnesota 55455, USA. EP - 3546 TI - Mechanisms of microtubule-based kinetochore positioning in the yeast metaphase spindle. VL - 84 SP - 3529 KW - kinetochore KW - positioning KW - yeast AU - Sprague, BL AU - Pearson, CG AU - Maddox, PS AU - Bloom, KS AU - Salmon, ED AU - Odde, DJ PY - 2003/06// UR - http://view.ncbi.nlm.nih.gov/pubmed/12770865 ER -