Contributions of protein aggregation to gene regulation and phenotypic diversity

摘要 蛋白质的聚集与人类疾病密切相关，包括数十种 家族性和与年龄相关的疾病共同构成对 我们老龄化的人口。然而，最近的发现表明，蛋白质聚集也可以 具有广泛的结构和监管职能。的广度和普及性 这种非病理性聚集在很大程度上是未被探索的。芽殖酵母中的酵母菌 Cerevisiae，多种内在无序蛋白(IDPs)，包括转录因子，RNA- 结合蛋白和激酶在生理条件下有聚集的趋势。 一种这样的蛋白质，转录因子mot3，是已知的少数几种蛋白质之一。 形成自我繁殖的聚集体，作为以蛋白质为基础的遗传元素，或称普恩。 通过切换到聚集状态和从聚集状态，mot3扩大了表型的范围 可供克隆酵母种群使用。这项工作的目标是1)开发一种量化的 国内流离失所者细胞内聚集的流式细胞仪报告，2)调查后果 关于国内流离失所者监管活动的聚合，以及3)测试这些聚合是否 蛋白质，尤其是mot3，是一个适应性的生物过程。这项工作将推动我们的 理解非病理性蛋白质聚集，同时建立面向未来的平台 活细胞中功能聚集和疾病相关聚集的询问。

ABSTRACT The aggregation of proteins is deeply associated with human diseases, including dozens of familial and age-associated disorders that together comprise a major emerging health threat to our aging populace. However, recent discoveries indicate that protein aggregation can also have a wide range of structural and regulatory functions. The breadth and pervasiveness of such non-pathological aggregation is largely unexplored. In the budding yeast, Saccharomyces cerevisiae, multiple intrinsically disordered proteins (IDPs), including transcription factors, RNA- binding proteins, and kinases, have a tendency to aggregate under physiological conditions. One such protein, the transcription factor Mot3, is one of only a handful of proteins known to form self-propagating aggregates that act as protein-based elements of inheritance, or prions. By switching to and from its aggregated state, Mot3 broadens the range of phenotypes accessible to clonal yeast populations. The goals of this work are to 1) develop a quantitative flow cytometric reporter for intracellular aggregation by IDPs, 2) investigate the consequences of aggregation on the regulatory activities of IDPs, and 3) test whether aggregation by these proteins, and by Mot3 in particular, is an adaptive biological process. This work will advance our understanding of non-pathological protein aggregation, while establishing a platform for future interrogations of both functional and disease-associated aggregation in living cells.