Mechanism of Spindle Pole Body Duplication and Separation in Yeast Meiosis

酵母减数分裂纺锤体复制与分离的机制

• 批准号: 1121771
• 负责人: Hong-Guo Yu
• 金额: $ 65.49万
• 依托单位: Florida State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-11-01 至 2016-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1121771&HistoricalAwards=false
• 关键词: Mechanism; Spindle; Pole; Body; Duplication

Intellectual meritThe yeast centrosome, also called the spindle pole body, organizes microtubules required for a variety of cellular processes that include cell division. During meiosis, the centrosome is duplicated when chromosomes replicate. Duplicated centrosomes separate to form a bipolar spindle required for homolog separation during the first division of meiosis. Centrosomes duplicate again in the absence of chromosome replication to establish two spindles for sister chromatid separation in the second meiotic division. Coordination of centrosome duplication and separation with the two chromosome segregation cycles in meiosis is fundamental to maintain genome integrity in eukaryotes. The goal of this project is to determine the molecular mechanism by which the Aurora kinase Ipl1 and the Polo-like kinase Cdc5 act antagonistically to regulate centrosome duplication and separation. Using creative yeast genetics in combination with live-cell microscopy and biochemical methods, the temporal and spatial requirements of Ipl1 and Cdc5 in centrosome duplication will be determined. In addition, their phosphorylated substrates will be identified by a novel phosphoproteome approach and characterized for their biochemical activities in regulation of centrosome dynamics. These studies will provide a mechanistic view of protein phosphorylation mediated by Ipl1 and Cdc5 in yeast centrosome duplication and have general implications for understanding the mechanism of centrosome dynamics in animals.Broader impactsThis project will train graduate and undergraduate students in genetics and cell biology, and will also provide unique opportunities for underrepresented and minority students to design and conduct goal-oriented experiments in their classroom. Students enrolled in the Experimental Biology course, which the PI teaches once a year, will tag centrosome components with the green fluorescent protein and screen for genetic abnormalities of the centrosome in yeast. This research will provide molecular insights into the regulation of centrosome duplication and contribute to society by educating those who will be part of our future work force as scholars and scientists.

智能优点酵母中心体，也被称为纺锤体极体，组织包括细胞分裂在内的各种细胞过程所需的微管。在减数分裂过程中，中心体在染色体复制时复制。在减数分裂的第一次分裂中，重复的中心体分开，形成同源分离所需的两极纺锤体。中心体在没有染色体复制的情况下再次复制，以建立两个纺锤体，用于第二次减数分裂中姐妹染色单体的分离。中心体复制和分离与减数分裂中的两个染色体分离循环的协调是维持真核生物基因组完整性的基础。该项目的目标是确定Aurora激酶Ipl1和Polo样激酶CDC5相互拮抗地作用于调节中心体复制和分离的分子机制。利用创造性的酵母遗传学，结合活细胞显微镜和生化方法，将确定中心体复制中Ipl1和CDc5的时间和空间需求。此外，它们的磷酸化底物将通过一种新的磷酸蛋白质组方法进行鉴定，并表征它们在调节中心体动力学方面的生化活性。这些研究将提供酵母中心体复制中Ipl1和CDC5介导的蛋白质磷酸化的机制观点，并对理解动物中心体动力学的机制具有普遍意义。Broader Impact本项目将培训研究生和本科生遗传学和细胞生物学，并将为代表不足的学生和少数族裔学生提供独特的机会，在他们的课堂上设计和进行目标导向的实验。PI每年教授一次实验生物学课程，注册的学生将用绿色荧光蛋白标记中心体组件，并筛查酵母中中心体的遗传异常。这项研究将为中心体复制的调节提供分子洞察力，并通过教育那些将成为我们未来工作大军的学者和科学家来为社会做出贡献。