Career: Molecular and Genetic Analysis of the Yeast EBP2 Gene

该研究项目将侧重于使用分子技术来表征一个新的蛋白质家族（Ebp2蛋白质）。Ebp2蛋白是重要的，因为它们是真核细胞分裂所必需的。 由于酵母中可用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，因此来自S.将研究酿酒。 初步分析表明，酵母Ebp2p是一种必需的核仁蛋白，并且ebp2突变体遭受与rRNA代谢和染色体分离相关的缺陷。为了更好地了解Ebp2p在细胞代谢中的作用，将创建和表征新的ebp2突变体。随机和定点诱变将用于改变Ebp2蛋白内的关键残基，并将研究这些突变以确定它们对核仁代谢和核分裂的影响。抑制基因分析和合成致死性的遗传学方法将用于鉴定与Ebp2p相互作用的蛋白质。为了进一步确定Ebp2p在核代谢中的作用，将使用显微镜分析来探测野生型和突变型ebp2细胞中的核结构。还将研究Ebp2蛋白，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。由于分析酵母中的一种新蛋白质（Ebp2p）提供了一个很好的机会，让学生参与基础研究，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能力项目合作，开办一个生物技术讲习班，对高中和初中教师进行基础分子生物学和遗传学实验方面的培训。这些研讨会将使用酵母和细菌作为模式生物，他们将强调动手的方法来教授高中和初中生遗传学的基本概念。

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.