The Role of Stu1 in Mitotic Spindle Stability

Stu1 在有丝分裂纺锤体稳定性中的作用

• 批准号: 7254246
• 负责人: IRENE Alexandra AMARO
• 金额: $ 3.62万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7254246
• 关键词: Affinity Chromatography; Aneuploid Cells; Antibodies; Biochemical; Biochemistry; Cell Cycle; Cell division; Class; Complex; Depth; Eukaryota; Eukaryotic Cell; Genetic; Genetic Materials; Genetic Screening; Hereditary Disease; Homologous Gene; Immunofluorescence Immunologic; In Vitro; Kinetochores; Lead; Localized; Maintenance; Mass Spectrum Analysis; Microtubule-Associated Proteins; Microtubules; Mitotic spindle; Modeling; Motor; Open Reading Frames; Protein Family; Proteins; Role; Saccharomyces cerevisiae Proteins; Structure; System; Testing; Xenopus; Xenopus Proteins; Xenopus laevis; Yeast Model System; Yeasts; cDNA Library; cancer genetics; egg; genetic analysis; insight; novel; segregation

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.

描述（由申请人提供）：细胞分裂过程中遗传物质的正确分离至关重要，并且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。正如在癌症和遗传疾病中观察到的那样，错误分离产生的非整倍体细胞会遭受有害影响并降低生存能力。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区，对于正确的纺锤体结构和活力至关重要。然而，与 Stu1 相互作用的蛋白质以及 Stu1p 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) 和质谱法鉴定 Stu1 关联蛋白将有助于了解纺锤体中 Stu1 的背景：酵母是一个有价值的遗传系统，但深入的生物化学是有限的。非洲爪蟾卵提取物是研究纺锤体形成和微管动力学的强大体外系统。使用从编码 Stu1 同源物的 cDNA 文库中鉴定出的非洲爪蟾蛋白抗体对提取物进行免疫定位和免疫耗竭，将允许在双极纺锤体形成和微管动力学中分析此类蛋白质。