Variable penetrance of Nab3 granule accumulation quantified by a new tool for high-throughput single-cell granule analysis.
Variable penetrance of Nab3 granule accumulation quantified by a new tool for high-throughput single-cell granule analysis.
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DOI：
10.1007/s00294-022-01234-2
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发表时间：
2022-08
期刊：
影响因子：
2.5
通讯作者：
中科院分区：
文献类型：
--
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细胞蛋白重组成亚细胞区室，如RNA结合蛋白浓缩成细胞质应激颗粒和P体，是目前正在深入研究的公认的、广泛研究的生理过程。这方面的一个例子是诱导酵母Nab3转录终止因子从其泛核质分布重排到核周边的颗粒，以响应营养限制。最近在许多细胞类型中的研究表明，细胞核中的蛋白质凝聚对于转录起始、RNA加工和终止具有重要的功能。然而，人们对亚核隔室如何形成知之甚少。在这里，我们已经定量分析了这个动态过程中活酵母使用高通量计算工具和荧光显微镜。该分析揭示了Nab3颗粒积累在不同酵母菌株中的突变率不同。浓缩的单个颗粒由至少四分之一的核Nab 3形成，该核Nab 3来自核的其余部分。颗粒积累的水平呈负相关，在没有葡萄糖的生长缺陷。重要的是，外显率的一些变化的基础可归因于线粒体功能的缺陷。这种公开可用的计算工具提供了一个严格的，可重复的，和公正的检查Nab3颗粒的积累，应广泛适用于各种荧光图像。可以很容易地检查数千个活细胞，从而能够对显著性进行严格的统计验证。利用它，我们描述了一个新的功能，可诱导的亚核区室形成的RNA结合转录因子和颗粒生物发生的一个重要决定因素。
Reorganization of cellular proteins into subcellular compartments, such as the concentration of RNA-binding proteins into cytoplasmic stress granules and P-bodies, is a well-recognized, widely studied physiological process currently under intense investigation. One example of this is the induction of the yeast Nab3 transcription termination factor to rearrange from its pan-nucleoplasmic distribution to a granule at the nuclear periphery in response to nutrient limitation. Recent work in many cell types has shown that protein condensation in the nucleus is functionally important for transcription initiation, RNA processing, and termination. However, little is known about how subnuclear compartments form. Here, we have quantitatively analyzed this dynamic process in living yeast using a high-throughput computational tool and fluorescence microscopy. This analysis revealed that Nab3 granule accumulation varies in penetrance across yeast strains. A concentrated single granule is formed from at least a quarter of the nuclear Nab3 drawn from the rest of the nucleus. Levels of granule accumulation were inversely correlated with a growth defect in the absence of glucose. Importantly, the basis for some of the variation in penetrance was attributable to a defect in mitochondrial function. This publicly available computational tool provides a rigorous, reproducible, and unbiased examination of Nab3 granule accumulation that should be widely applicable to a variety of fluorescent images. Thousands of live cells can be readily examined enabling rigorous statistical verification of significance. With it, we describe a new feature of inducible subnuclear compartment formation for RNA-binding transcription factors and an important determinant of granule biogenesis.
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DOI：
10.1042/bcj20160499
发表时间：
2017-04-07
期刊：
The Biochemical journal
影响因子：
--
作者：
Harrison AF;Shorter J
通讯作者：
Shorter J
影响因子：
5.3
作者：
Jenks, M. Harley;O'Rourke, Thomas W.;Reines, Daniel
通讯作者：
Reines, Daniel
影响因子：
16
作者：
Lin Y;Protter DS;Rosen MK;Parker R
通讯作者：
Parker R
影响因子：
3.7
作者：
Lee S;Lim WA;Thorn KS
通讯作者：
Thorn KS
影响因子：
16
作者：
Kuehner, Jason N.;Brow, David A.
通讯作者：
Brow, David A.