Dynamics of yeast prion aggregates in single living cells
Dynamics of yeast prion aggregates in single living cells
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DOI：
10.1111/j.1365-2443.2006.01004.x
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发表时间：
2006-09-01
期刊：
影响因子：
2.1
通讯作者：
Taguchi, Hideki
中科院分区：
文献类型：
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作者：
Kawai-Noma, Shigeko;Ayano, Satoru;Taguchi, Hideki
朊病毒是一种繁殖蛋白，它是有序的蛋白质聚集体，其表型特征保留在改变的蛋白质构象中。为了了解活细胞中朊病毒聚集体的动态，我们使用片上单细胞培养系统和荧光相关光谱（FCS）直接监测聚集体的命运。单细胞成像显示，与GFP融合的酵母朊病毒Sup35的可见病灶在细胞生长过程中分散在整个细胞质中，但保留了朊病毒的表型。FCS显示，无论是否存在病灶，[PSI+]细胞都含有弥漫性低聚物，这些低聚物会传递给它们的子细胞。基于寡聚物的传播的单细胞观察提供了先前对朊病毒的体内和体外分析之间的联系，并阐明了蛋白质构象与表型之间的关系。
Prions are propagating proteins that are ordered protein aggregates, in which the phenotypic trait is retained in the altered protein conformers. To understand the dynamics of the prion aggregates in living cells, we directly monitored the fate of the aggregates using an on-chip single-cell cultivation system as well as fluorescence correlation spectroscopy (FCS). Single-cell imaging revealed that the visible foci of yeast prion Sup35 fused with GFP are dispersed throughout the cytoplasm during cell growth, but retain the prion phenotype. FCS showed that [PSI+] cells, irrespective of the presence of foci, contain diffuse oligomers, which are transmitted to their daughter cells. Single-cell observations of the oligomer-based transmission provide a link between previous in vivo and in vitro analyses of the prion and shed light on the relationship between the protein conformation and the phenotype.