Career: Molecular and Genetic Analysis of the Yeast EBP2 Gene

本研究项目将侧重于利用分子技术表征一个新的蛋白质家族（Ebp2蛋白）。eebp2蛋白是真核生物细胞分裂所必需的。由于酵母的分子和遗传技术为研究细胞生物学的基本方面提供了强有力的方法，因此将研究酿酒酵母的EBP2同源物。初步分析表明，酵母ebp2是一种必需的核仁蛋白，而ebp2突变体存在与rRNA代谢和染色体分离相关的缺陷。为了更好地了解ebp2在细胞代谢中的作用，新的ebp2突变体将被创建和表征。随机和定点突变将用于改变Ebp2蛋白内的关键残基，这些突变将被研究以确定它们对核仁代谢和核分裂的影响。抑制因子分析和合成致死率的遗传方法将用于鉴定与Ebp2p相互作用的蛋白质。为了进一步确定ebp2在核代谢中的作用，我们将使用显微镜分析来探测野生型和突变型ebp2细胞的核结构。Ebp2蛋白也将被研究，以确定它是否与染色质相关，以及它是否在核仁rDNA重复序列的染色体凝聚中起作用。由于酵母中一种新蛋白的分析（Ebp2p）为学生从事基础研究提供了一个很好的机会，这些研究将与学生的分子遗传学教育相协调。此外，还将与提高数学和科学水平项目合作，设立一个生物技术讲习班，培训高中和初中教师进行基本分子生物学和遗传学实验。这些工作坊将以酵母菌和细菌作为模式生物，并强调向高中和初中学生讲授遗传学基本概念的实践方法。

This research project will focus on using molecular techniques to characterize a new family of proteins (the Ebp2 proteins). The Ebp2 proteins are significant because they are required for eukaryotic cellular division. Since the molecular and genetic techniques available in yeast provide powerful approaches for studying basic aspects of cellular biology, the EBP2 homolog from S. cerevisiae will be studied. Preliminary analysis indicates that yeast Ebp2p is an essential nucleolar protein, and that ebp2 mutants suffer from defects related to rRNA metabolism and chromosome segregation. In order to gain a greater understanding of the role that Ebp2p plays in cellular metabolism, new ebp2 mutants will be created and characterized. Random and site-directed mutagenesis will be used to alter critical residues within the Ebp2 protein, and these mutations will be studied to determine the effects that they have on nucleolar metabolism and nuclear division. The genetic approaches of suppressor analysis and synthetic lethality will be used to identify proteins that interact with Ebp2p. To further define the role that Ebp2p plays in nuclear metabolism, microscopic analysis will be used to probe nuclear structure in wild type and mutant ebp2 cells. The Ebp2 protein will also be investigated to determine if it is associated with chromatin and whether it functions in chromosome condensation of the nucleolar rDNA repeat sequences. Since the analysis of a novel protein in yeast (Ebp2p) offers a great opportunity to engage students in basic research, these studies will be coordinated with the education of students in molecular genetics. Also, in collaboration with the Project to Increase Mastery of Mathematics and Science (PIMMS), a Biotechnology Workshop will be developed to train high school and middle school teachers in experiments in basic molecular biology and genetics. These workshops will use yeast and bacteria as model organisms, and they will emphasize hands-on approaches for teaching basic concepts of genetics to high school and middle school students.