Hsp104 binds to yeast Sup35 prion fiber but needs other factor(s) to sever it
Hsp104 binds to yeast Sup35 prion fiber but needs other factor(s) to sever it
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DOI：
10.1074/jbc.m408159200
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发表时间：
2004-12-10
影响因子：
4.8
通讯作者：
Yoshida, M
中科院分区：
文献类型：
--
作者：
Inou, Y;Taguchi, H;Yoshida, M
测试了 Hsp104 与由 Sup35NM（Sup35 的朊病毒诱导结构域）制成的酵母朊病毒纤维的相互作用。当荧光标记的 Hsp104 添加到预成型纤维中时，单根纤维沿纤维长度均匀地进行荧光装饰。然而，一种纤维与另一种纤维的荧光密度不同，表明存在 Sup35NM 纤维亚种。 Hsp104 延迟了单体 Sup35NM 形成纤维的时间进程。使用珠系纤维测试 Hsp104 介导的纤维断裂。与最近的报告（Shorter, J., and Lindquist, S. (2004) Science 304, 1793-1797）相比，单独使用 Hsp104 无法切断纤维。酵母细胞裂解物或缺乏 Hsp104 的细胞裂解物加 Hsp104 会导致 ATP 依赖性、盐酸胍敏感的纤维断裂。因此，在我们的实验设置中，需要 Hsp104 加上酵母细胞质中的其他因子来切断酵母朊病毒纤维。上述报告差异的原因尚不清楚，但可能是由于朊病毒纤维的构象亚种不同造成的。
The interaction of Hsp104 with yeast prion fibers made of Sup35NM, a prion-inducing domain of Sup35, was tested. When fluorescently labeled Hsp104 was added to the preformed fibers, individual fibers were fluorescently decorated uniformly along the fiber length. However, the density of fluorescence differed from one fiber to another, indicating the presence of subspecies of Sup35NM fibers. The time course of fiber formation from monomer Sup35NM was delayed by Hsp104. Hsp104-mediated fragmentation of fibers was tested using bead-tethered fibers. In contrast with the recent report (Shorter, J., and Lindquist, S. (2004) Science 304, 1793-1797), Hsp104 alone was unable to sever the fibers. Yeast cell lysate or the Hsp104-deficient cell lysate plus Hsp104 caused ATP-dependent, guanidine hydrochloride-sensitive fragmentation of the fibers. Thus, in our experimental setup, Hsp104 plus other factor(s) in the yeast cytosol are required for severing yeast prion fiber. The reason of discrepancy from the above report is unknown but is possibly caused by different conformational subspecies of prion fibers.