Chernoff Y O, Galkin A P, Lewitin E, Chernova T A, Newnam G P, Belenkiy S M
School of Biology, Georgia Institute of Technology, Atlanta, GA 30332-0230, USA.
Mol Microbiol. 2000 Feb;35(4):865-76. doi: 10.1046/j.1365-2958.2000.01761.x.
Saccharomyces cerevisiae prion [PSI ] is a self-propagating isoform of the eukaryotic release factor eRF3 (Sup35p). Sup35p consists of the evolutionary conserved release factor domain (Sup35C) and two evolutionary variable regions - Sup35N, which serves as a prion-forming domain in S. cerevisiae, and Sup35M. Here, we demonstrate that the prion form of Sup35p is not observed among industrial and natural strains of yeast. Moreover, the prion ([PSI + ]) state of the endogenous S. cerevisiae Sup35p cannot be transmitted to the next generations via heterologous Sup35p or Sup35NM, originating from the distantly related yeast species Pichia methanolica. This suggests the existence of a 'species barrier' in yeast prion conversion. However, the chimeric Sup35p, containing the Sup35NM region of Pichia, can be turned into a prion in S. cerevisiae by overproduction of the identical Pichia Sup35NM. Therefore, the prion-forming potential of Sup35NM is conserved in evolution. In the heterologous system, overproduction of Pichia Sup35p or Sup35NM induced formation of the prion form of S. cerevisiae Sup35p, albeit less efficiently than overproduction of the endogenous Sup35p. This implies that prion induction by protein overproduction does not require strict correspondence of the 'inducer' and 'inducee' sequences, and can overcome the 'species barrier'.
酿酒酵母朊病毒[PSI+]是真核释放因子eRF3(Sup35p)的一种自我传播异构体。Sup35p由进化保守的释放因子结构域(Sup35C)和两个进化可变区域组成——Sup35N,它在酿酒酵母中作为朊病毒形成结构域,以及Sup35M。在此,我们证明在工业酵母菌株和天然酵母菌株中未观察到Sup35p的朊病毒形式。此外,酿酒酵母内源性Sup35p的朊病毒([PSI+])状态不能通过源自远缘酵母物种甲醇毕赤酵母的异源Sup35p或Sup35NM传递给下一代。这表明酵母朊病毒转化中存在“物种屏障”。然而,含有甲醇毕赤酵母Sup35NM区域的嵌合Sup35p,通过过量表达相同的甲醇毕赤酵母Sup35NM,可在酿酒酵母中转变为朊病毒。因此,Sup35NM的朊病毒形成潜力在进化中是保守的。在异源系统中,过量表达甲醇毕赤酵母Sup35p或Sup35NM可诱导酿酒酵母Sup35p形成朊病毒形式,尽管效率低于过量表达内源性Sup35p。这意味着蛋白质过量表达诱导朊病毒形成并不需要“诱导物”和“被诱导物”序列严格对应,并且可以克服“物种屏障”。