Studies of Telomere Structure Using Yeast Model Systems

在过去的三年里，在夸美纽斯大学卢博米尔·托马斯卡博士的实验室工作 斯洛伐克共和国得到了FIRCA奖(两个5654)的支持，父母授予GM31819博士。 北卡罗来纳大学的杰克·格里菲斯。FIRCA的支持产生了14种出版物和10种 托马斯卡实验室过去3年的会议摘要和两篇备受瞩目的论文 发表在《生物化学杂志》上。这些研究为调查奠定了基础 端粒结构被美国和美国的研究人员视为非常重要的 在国际上。这项工作主要是在夸美纽斯大学完成的，每年有1到3个月的访问 托马斯卡博士和他的学生给北卡罗来纳大学。该奖项的所有资金都已捐给了他的实验室。在AIM I中， 酿酒酵母微染色体系统的开发将允许分离天然的 用于电子显微镜(EM)检查的微小染色体提供了一种重要的新工具，将 直接惠及父母拨款的目标和格里菲斯实验室的目标。这 托马斯卡实验室将使用微型染色体系统继续检查 酵母端粒的结构。在AIM II，也在FIRCA奖下启动的工作将继续 利用S.pombe系统在体外重建酵母端粒。在过去的资助期，端粒 结合蛋白Taz1被证明在模型庞贝链霉菌端粒末端形成环。这就是现在的情况 美国能够理解Taz1蛋白如何与POT1和其他端粒结合蛋白相互作用 形成一个天然的端粒。NIH的母公司拨款支持了检查方法的发展 DNA-蛋白质复合体，并将这些方法转移到其他实验室。这项技术 转移现在已经开始，托马斯卡博士的学生学习这些EM方法。至关重要的是，我们 继续这一卓有成效的国际合作。

Over the past three years work in the laboratory of Dr. Lubomir Tomaska at Comenius University in the Slovak Republic has been supported by a FIRCA award (TWO 5654) under the parent grant GM31819 to Dr. Jack Griffith at the University of North Carolina. The FIRCA support has resulted in 14 publications and 10 meeting abstracts from the Tomaska laboratory in the past 3 years with two high profile papers just published in TheJournalof Biological Chemistry. These studies have laid the foundation for investigations of telomere structure which are being seen as very significant by investigators both in the US and internationally. The work has been done primarily at Comenius University with 1to 3 month visits each year by Dr. Tomaska and his students to UNC. All funds from the award have gone to his laboratory. In Aim I, the development of the yeast S. cerevisciae minichromosome system will allow the isolation of native minichromosomes for examination by electron microscopy (EM) providing an important new tool that will directly benefit the goals of the parent grant and the objectives of the Griffith laboratory. This minichromosome system will be used by the Tomaska laboratory to continue their examination of the architecture of yeast telomeres. In Aim II, work also initiated under the FIRCA award will continue on the reconstitution of a yeast telosome in vitro using the S. pombe system. In the past funding period the telomere binding protein, Taz1 was shown to form loops at the ends of model S. pombe telomeres. This now places us in a position to understand how Taz1 protein interacts with Pot1 and other telomere binding proteins to form a native telosome. The parent NIH grant has supported the development of methods for examining DNA-protein complexes by EM, and transferring these methods to other laboratories. This technology transfer has now begun with Dr. Tomaska's students learning these EM methods. It is critical that we continue this highly productive international collaboration.