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，包括数十种疾病 家族性疾病和与年龄相关的疾病共同构成了新出现的主要健康威胁 我们的老龄化人口。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。的广度和普遍性 这种非病理性聚集在很大程度上尚未被探索。在芽殖酵母中，酿酒酵母 酿酒酵母，多种内在无序蛋白（IDP），包括转录因子、RNA- 结合蛋白和激酶在生理条件下有聚集的倾向。 其中一种蛋白质，转录因子 Mot3，是已知的少数蛋白质之一。 形成自我繁殖的聚集体，充当基于蛋白质的遗传元件或朊病毒。 通过在聚合状态之间切换，Mot3 拓宽了表型范围 可用于克隆酵母群体。这项工作的目标是 1）开发一个定量的 IDP 细胞内聚集的流式细胞报告仪，2) 研究后果 国内流离失所者监管活动的汇总，以及 3) 测试这些汇总是否 蛋白质，尤其是 Mot3，是一种适应性生物过程。这项工作将推动我们 了解非病理性蛋白质聚集，同时为未来建立平台 对活细胞中功能性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.