Biochemical and Structural Studies of Gamma-Tubulin in Arabidopsis Thaliana

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9808480OakleyBiochemical and Structural Studies of Gamma-Tubulin in ArabidopsisAbstractMicrotubule arrays are essential for cell division and morphogenesis in plant cells, but how these arrays assemble and rearrange is largely unknown. In animal and fungal cells a protein called gamma-tubulin is important for microtubule assembly. In animal cells gamma-tubulin forms large ring-shaped complexes that nucleate microtubule assembly and remain attached to the microtubule ends. In plant cells gamma-tubulin is also present, but how it associates with microtubules is unclear. Ring complexes have not yet been identified, and whether or how gamma-tubulin might control microtubule assembly is unknown. The overall goal of this project is to clarify the functions of gamma-tubulin in plant cells. The first specific goal is to determine if gamma-tubulin forms ring complexes in cells of Arabidopsis thaliana and if they do form, to purify them and determine their ability to nucleate microtubule assembly. The second specific goal is to determine whether purified Arabidopsis gamma-tubulin binds in vitro at the ends of microtubules or along their sides. Evidence for ring complexes and nucleation of assembly by gamma-tubulin will support an important role of gamma-tubulin in formation of microtubule arrays in plant cells in vivo. If gamma-tubulin behaves differently in plant cells than in animal cells, the studies will establish a good starting point to determine the actual functions of this protein in plant cells.