The Role of Stu1 in Mitotic Spindle Stability

描述（由申请方提供）：细胞分裂期间遗传物质的适当分离至关重要，并取决于由微管和相关蛋白组成的双极有丝分裂纺锤体的形成和稳定性。如在癌症和遗传疾病中观察到的，由错误分离产生的非整倍体细胞遭受有害影响和活力降低。酿酒酵母蛋白Stulp与微管相互作用，定位于有丝分裂纺锤体中间区，并且对于适当的纺锤体结构和活力是必需的。然而，与Stu 1相互作用的蛋白质以及Stu 1p在有丝分裂纺锤体形成和维持中的作用尚不清楚。使用串联亲和纯化（TAP）和质谱鉴定Stu 1相关蛋白将有助于理解Stu 1在纺锤体中的背景：酵母是一种有价值的遗传系统，但在深度生物化学方面是有限的。非洲爪蟾卵提取物是研究纺锤体形成和微管动力学的有效体外系统。免疫定位和免疫耗竭的提取物，使用抗体对非洲爪蟾蛋白的cDNA文库编码的Stu1同源物将允许这类蛋白质的双极纺锤体的形成和微管动力学的分析。

DESCRIPTION (provided by applicant): Proper segregation of genetic material during cell division is critical and depends on the formation and stability of the bipolar mitotic spindle composed of microtubules and associated proteins. Aneuploid cells resulting from mis-segregation suffer detrimental effects and reduced viability as observed in cancers and genetic diseases. The Saccharomyces cerevisiae protein Stulp interacts with microtubules, localizes to the mitotic spindle midzone region, and is essential for proper spindle structure and viability. However, proteins that interact with Stu1 and the role of Stu1p in formation and maintenance of the mitotic spindle is not understood. Identification of Stu1 associating proteins using Tandem Affinity Purification (TAP) and mass spectroscopy will aid in understanding the context of Stu1 in the spindle: Yeast is a valuable genetic system, but in depth biochemistry is limited. Xenopus laevis egg extracts are a powerful in vitro system for studying spindle formation and microtubule dynamics. Immunolocalization and immunodepletion of extracts using antibodies against a Xenopus protein identified from cDNA libraries encoding a Stu1 homologue will allow analysis of this class of proteins in bipolar spindle formation and microtubule dynamics.