CAREER:Investigating Heterochromatin Assembly through Histone Deacetylases

细胞基因组被“包装”在细胞核内的方式直接影响基因组的稳定性和dna模板化过程，如细胞分裂所必需的基因表达和复制。高度浓缩的DNA结构对这些过程有抑制作用，它们的形成是调控机制的关键部分。尽管具有重要的生物学作用，但人们对高度浓缩的DNA区域（称为异染色质）是如何形成的知之甚少。这个职业项目将阐明和比较不同功能异染色质结构域形成和繁殖所必需的分子途径。Wiley博士和一组本科生将利用亲热四膜虫（Tetrahymena thermophila）的独特特征，通过确定其空间和时间定位以及分析突变细胞，专门探索组蛋白去乙酰化酶（HDAC）酶在异染色质形成中的作用。使用hdac作为分子手柄，学生将使用标准的分子技术分离和识别与hdac结合在复合物中的蛋白质，并进一步分析这些蛋白质作为途径的候选成分。该项目的设计将显著增加本科生参与原创研究的人数。在当地，这项工作将作为大二学生的新实验课程的基础，以及许多指导学生的研究项目。为了将研究机会扩展到其他院校的本科生，相关的协议和方法将打包并传播为课堂实验活动，以便于整合到现有的课程中。来自所有相关机构的学生研究人员将通过一个与斯坦福大学四膜虫基因组数据库相连的互动网站/数据库，与更广泛的科学界传播和讨论他们的结果。这个项目直接有助于我们理解分子生物学中的一个核心问题，这个问题是生命必不可少的过程的基础——基因组如何描绘和调节控制细胞特征的信息。它将增强我们对基因组动力学的理解，对最近测序的基因组的注释做出重要贡献，也将为更大的科学界进一步的基因组研究提供有价值的试剂。通过这一多机构倡议，许多学生团队将参与到合作研究网络中，并成为基于原始研究兴奋的强大科学学习社区的一部分，所有这些都是在他们做出职业决定的重要时刻。这种经历应该鼓励学生坚持他们的科学教育，并特别支持历史上代表性不足的群体。增加所有学生接触科学的机会，对于确保未来的技术进步和持续的经济活力至关重要，而我们的高水平生活和其他国家的发展都依赖于这些技术进步和经济活力。

The way that the genome of a cell is "packaged" inside the nucleus directly impacts genome stability and DNA-templated processes such as gene expression and replication necessary for cell division. Highly condensed DNA structures are inhibitory to such processes, and their formation is a critical part of regulatory mechanisms. Despite the important biological roles, little is known about how regions of highly condensed DNA, called heterochromatin, form. This CAREER project will elucidate and compare the molecular pathways necessary for the formation and propagation of functionally distinct heterochromatin domains. Exploiting unique features of the ciliate model Tetrahymena thermophila, Dr. Wiley and a team of undergraduate students will specifically explore the role of histone deacetylase (HDAC) enzymes in heterochromatin formation by determining their spatial and temporal localization and by analyzing mutant cells. Using HDACs as molecular handles, students will employ standard molecular techniques to isolate and identify proteins that bind to them in complexes and further analyze these proteins as candidate components of the pathways. The project's design will significantly increase the number of undergraduates participating in original research. Locally, the work will serve as the foundation of a new laboratory course for sophomore level students, and for numerous mentored student research projects. To extend the research opportunities to undergraduate students at other institutions, the relevant protocols and methods will be packaged and disseminated as classroom laboratory activities for easy integration into existing curricula. Student researchers from all institutions involved will disseminate and discuss their results with the broader scientific community through an interactive website/database linked to the Tetrahymena Genome Database at Stanford University. This project contributes directly to our understanding of a central question in molecular biology that underlies processes essential for life -- how genomes portray and regulate the information that governs cell characteristics. It will enhance our understanding of genome dynamics, make important contributions to the annotation of a recently sequenced genome, and will also provide to the larger scientific community, valuable reagents for further genome research. Through this multi-institutional initiative, many teams of students will participate in the collaborative research network and be part of a powerful scientific learning community based on the excitement of original research, all at an important time when they are making career decisions. This experience should encourage students to persist in their science education and be particularly supportive of historically underrepresented groups. Increasing the accessibility of science for all students is critical to ensuring future technological advancements and continued economic vitality upon which our high standard of living and the development of other nations depend.