Prion-based regulation of RNA-modifying enzyme activities

项目摘要/摘要 Pron蛋白在生物学上是臭名昭著的，因为它会导致致命的、传染性的脑部疾病。 然而，除了疾病之外，普恩病毒完全改变了我们对遗传的理解，而且 与它们的名声相反，它们不仅是致病因子。相反，他们是 在生命王国中发现，是一种表观遗传机制，影响整体 细胞过程，也产生对生物有益的特征。在这份提案中，我们 重点关注调节RNA的普恩，特别是RNA的启动子构象- 修饰酶。这些高度保守的酶催化了许多化学物质 对RNA的修饰对其结构和功能至关重要。我们的目标是 研究是为了了解这些酶的普恩形式如何改变它们的活动。 在RNA上，产生基因表达的变化，最终导致长期的， 可遗传的生长表型，如对环境胁迫的抗性。最近的一次 我们已经详细说明这样的例子是伪尿嘧啶合成酶的原生形式， 影响细胞增殖、衰老和蛋白质合成。这项工作使我们得以 研究其他RNA修饰酶具有类似普里恩行为的能力， 现在我们有了几个新的研究例子。在这份提案中，我们将调查 当使RNA修饰的酶采用朊病毒时，RNA修饰是否发生变化- 就像构象一样，这些变化对基因表达的影响，对 酶催化，传染性普恩蛋白的结构，以及普恩是否- 我们在发芽酵母中观察到的基于行为的行为在人类细胞中是保守的。我们 结合遗传学、高通量测序、 经典的生化方法和荧光显微镜。总之，这项工作的目的是 揭示表观遗传的、基于PrP的RNA调控的分子细节 细胞的生长和存活。

Project Summary/Abstract Prion proteins are notorious in biology for causing deadly, infectious brain diseases. Beyond disease, however, prions completely altered our understanding of heredity, and contrary to their reputation, they are not only disease-causing agents. Instead, they are found across kingdoms of life, and are an epigenetic mechanism that affects a panoply of cellular processes, also producing biologically beneficial traits. In this proposal we focus on prions that regulate RNA, specifically, prionogenic conformations of RNA- modifying enzymes. These highly conserved enzymes catalyze numerous chemical modifications on RNA that are critical for their structure and function. The goal of our research is to understand how prion-like forms of these enzymes may alter their activities on RNA, producing changes to gene expression, and ultimately, leading to long-lasting, heritable growth phenotypes, such as resistance to environmental stress. One recent such example that we have detailed is a prionogenic form of a pseudouridine synthase, which affects cell proliferation, aging, and protein synthesis. This work led us to investigate the capacity for other RNA-modifying enzymes to have prion-like behavior, and we now have several new examples for study. In this proposal, we will investigate whether RNA modifications are changed when the enzymes that make them adopt prion- like conformations, the effects these changes have on gene expression, the role for enzyme catalysis, the structure of the infectious prion protein, and whether the prion- based behavior that we have observed in budding yeast is conserved in human cells. We approach these questions using a combination of genetics, high-throughput sequencing, classical biochemical methods, and fluorescence microscopy. In summary, this work aims to uncover the molecular details of epigenetic, prion-based regulation of RNA that affects cell growth and survival.