Sequence Requirements for Protein-Based Genetic Elements

生物学中最基本的问题之一是生物体如何存储和传递遗传信息。表观遗传学不断发展的领域中的研究表明，遗传信息并不总是编码在生物体的DNA序列中。酵母普里恩的发现表明，蛋白质结构的变化可以用来传递遗传信息。酵母蛋白的形成涉及到将可溶性蛋白质转化为不溶的淀粉样蛋白。淀粉样蛋白可以稳定地从母细胞传递给子细胞，允许编码的表观遗传信息作为基于蛋白质的遗传元件传递。这个项目检验了这样的结构转换代表了真核生物细胞调控和遗传的常见表观遗传机制的假设。将使用体外和体内分析相结合的方法来确定允许某些蛋白质经历类病毒结构转换的序列特征。然后，这些信息将被用来识别其活性受类病毒结构转换调控的新蛋白质。综上所述，这些研究将促进我们对这种结构转换在真核调控和遗传中的作用的理解。广泛的影响：除了智力上的优点，这个项目还有三个主要领域的更广泛的影响。首先，大部分研究将在一门新的实验室课程的背景下进行，该课程被设计为独立研究和传统实验室课程的混合体。本课程将允许学生产生新的，可发表的研究，但在一个足够有组织的环境中，允许培训大量的学生。该课程旨在提供生物化学、分子生物学、遗传学和生物信息学方面的多样化跨学科培训经验。其次，由于酵母是安全和廉价的，它们为向年轻人传授生物化学和遗传学的基本概念提供了一个极好的系统。因此，这项研究的某些方面将用于一个名为“生物化学是小学”的小学推广项目。这个项目通过激动人心的动手活动，向学生介绍生物化学的科学过程和基本概念。最后，这项工作涉及开发定量监测Pron动力学的新技术，这将应用于未来对调节Prion的研究和淀粉样纤维的工业应用。

One of the most fundamental questions in biology is how organisms store and pass on genetic information. Research in the growing field of epigenetics has demonstrated that genetic information is not always encoded within an organism's DNA sequence. The discovery of yeast prions showed that changes in protein structure can be used to transmit genetic information. The formation of yeast prions involves the conversion of soluble proteins into an insoluble amyloid form. The amyloid form can be stably passed from mother to daughter cell, allowing the encoded epigenetic information to be passed on as a protein-based genetic element. This project examines the hypothesis that such structural conversions represent a common epigenetic mechanism of cellular regulation and inheritance in eukaryotes. A combination of in vitro and in vivo assays will be employed to define the sequence features that allow certain proteins to undergo prion-like structural conversions. This information will then be used to identify novel proteins whose activities are regulated by prion-like structural conversions. Taken together, these studies will advance our understanding of the role of such structural conversions in eukaryotic regulation and inheritance.Broader Impacts: In addition to the intellectual merit, this project has three major areas of broader impact. First, much of the research will be conducted in the context of a novel laboratory course, which is designed to be a hybrid of independent research and traditional laboratory classes. This course will allow students to generate new, publishable research, but in a sufficiently structured setting to allow for training of a large number of students. The course is designed to offer a diverse interdisciplinary training experience in biochemistry, molecular biology, genetics and bioinformatics. Second, as yeast are safe and inexpensive to work with, they provide an excellent system for teaching basic concepts of biochemistry and genetics to younger ages. Therefore, aspects of the research will be used in an elementary school outreach project called "Biochemistry is Elementary." This project introduces students to the scientific process and to basic concepts in biochemistry using exciting, hands-on activities. Finally, this work involves the development of novel technologies for the quantitative monitoring of prion dynamics, which will have applications for future research into regulatory prions and for industrial applications of amyloid fibrils.