The yeast prion [URE3] can be greatly induced by a functional mutated URE2 allele
The yeast prion [URE3] can be greatly induced by a functional mutated URE2 allele
复制标题
DOI：
10.1093/emboj/19.13.3215
复制
发表时间：
2000-07-03
期刊：
影响因子：
11.4
通讯作者：
Cullin, C
中科院分区：
文献类型：
--
作者：
Fernandez-Bellot, E;Guillemet, E;Cullin, C
酵母的非孟德尔元件[URE 3]被认为是Ure 2蛋白的朊病毒形式。[URE 3]表型在单倍体酵母菌株中以10（-5）的频率发生，是可逆的，并且其频率通过过表达URE 2基因而增加。我们创造了一种新的Ure 2蛋白突变体，称为H2 p，它导致[URE 3]发生率增加1000倍。迄今为止，只有Ure 2 p的各种C-末端截短突变体的过表达才能达到相当的水平。因此，h2等位基因是第一个表征的URE 2等位基因，其在低水平表达时诱导朊病毒形成。通过改组突变和野生型结构域的URE 2，我们还创建了第一个突变的Ure 2蛋白，是功能性的，并诱导朊病毒的形成。我们证明，URE 2的结构域协同功能顺式诱导[URE 3]的形成，这突出了分子内相互作用的重要性Ure 2 p折叠。此外，我们显示使用绿色荧光蛋白（GFP）融合蛋白的h2等位基因表现出许多丝状结构，不产生的野生型蛋白。
The non-Mendelian element [URE3] of yeast is considered to be a prion form of the Ure2 protein. The [URE3] phenotype occurs at a frequency of 10(-5) in haploid yeast strains, is reversible, and its frequency is increased by overexpressing the URE2 gene. We created a new mutant of the Ure2 protein, called H2p, which results in a 1000-fold increase in the rate of [URE3] occurrence. To date, only the overexpression of various C-terminal truncated mutants of Ure2p gives rise to a comparable level. The h2 allele is, thus, the first characterized URE2 allele that induces prion formation when expressed at a low level. By shuffling mutated and wild-type domains of URE2, we also created the first mutant Ure2 protein that is functional and induces prion formation, We demonstrate that the domains of URE2 function synergistically in cis to induce [URE3] formation, which highlights the importance of intramolecular interactions in Ure2p folding. Additionally, we show using a green fluorescent protein (GFP) fusion protein that the h2 allele exhibits numerous filiform structures that are not generated by the wild-type protein.