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Drosophila EB1 Is Important for Proper Assembly and Positioning of the Mitotic Spindle

Steve Rogers and Ron Vale

Over the last few years, it has become evident that there is a growing class of microtubule-associated proteins (MAPs) that localize to growing MT plus ends in vivo. This localization is significant as it allows these proteins to modulate microtubule dynamics, mediate interactions between the microtubule tip and other cellular structures, and localize other 'passenger' proteins to the microtubule plus end. The EB1 family of proteins are one such class of plus end-binding MAP. First identified as a binding partner for the human adenomatous polyposis coli (APC) tumor suppressor protein, the EB1s have since been shown to be evolutionarily conserved from yeast to mammals. We have been studying the cellular functions of EB1 using cultured Drosophila Schneider (S2) cells as a model system. We find that, as in other systems, fly EB1 localizes to the tips of microtubules and to the centrosomes and spindle poles throughout the cell cycle. The use of RNAi to generate EB1 loss-of-function phenotypes revealed that interphase microtubule organization appeared normal. However, EB1-deficient mitotic cells exhibit an array of spindle defects, along with an elevated mitotic index. In addition, mitotic spindles, which usually self- center within these cells, often misposition away from the cell center. We also examined the contribution of EB1 to microtubule dynamics in vivo using newly-developed methods in fly cells.

ASCB 2001

 

last updated 1/7/02


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