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首页> 外文期刊>Molecular and Cellular Biology >Induction of Distinct [URE3] Yeast Prion Strains
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Induction of Distinct [URE3] Yeast Prion Strains

机译：[URE3]酵母Pri病毒菌株的诱导

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[URE3] is a non-Mendelian genetic element inSaccharomyces cerevisiae, which is caused by a prion-like, autocatalytic conversion of the Ure2 protein (Ure2p) into an inactive form. The presence of [URE3] allows yeast cells to take up ureidosuccinic acid in the presence of ammonia. This phenotype can be used to select for the prion state. We have developed a novel reporter, in which the ADE2 gene is controlled by the DAL5 regulatory region, which allows monitoring of Ure2p function by a colony color phenotype. Using this reporter, we observed induction of different [URE3] prion variants (“strains”) following overexpression of the N-terminal Ure2p prion domain (UPD) or full-length Ure2p. Full-length Ure2p induced two types of [URE3]: type A corresponds to conventional [URE3], whereas the novel type B variant is characterized by relatively high residual Ure2p activity and efficient curing by coexpression of low amounts of a UPD-green fluorescent protein fusion protein. Overexpression of UPD induced type B [URE3] but not type A. Both type A and B [URE3] strains, as well as weak and strong isolates of type A, were shown to stably maintain different prion strain characteristics. We suggest that these strain variants result from different modes of aggregation of similar Ure2p monomers. We also demonstrate a procedure to counterselect against the [URE3] state.
机译：[ URE3 ]是酿酒酵母中的非孟德尔遗传元素，它是由a病毒样Ure2蛋白（Ure2p）自催化转化为非活性形式引起的。 [ URE3 ]的存在可使酵母细胞在氨存在下吸收尿素琥珀酸。该表型可用于选择病毒状态。我们开发了一种新型的报告基因，其中 ADE2 基因受 DAL5 调控区域控制，该区域可通过菌落颜色表型监测Ure2p功能。使用该报告基因，我们观察到在N端Ure2p ion病毒结构域（UPD）或全长Ure2p过度表达后诱导了不同的[ URE3 ] ion病毒变体（“菌株”）。全长Ure2p诱导两种类型的[ URE3 ]：A型对应于常规的[ URE3 ]，而新型B型变体的特征是相对较高的残留Ure2p活性和通过共表达少量UPD-绿色荧光蛋白融合蛋白可实现高效固化。 UPD的过度表达诱导了B型[ URE3 ]而不是A型。A型和B型[ URE3 ]菌株以及A型的弱和强分离株都是可以稳定地维持不同的病毒应变特性。我们建议这些菌株变异是由相似的Ure2p单体的不同聚集方式引起的。我们还演示了针对[ URE3 ]状态进行反选择的过程。

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