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首页> 外文期刊>Molecular and Cellular Biology >Upf1p, Nmd2p, and Upf3p are interacting components of the yeast nonsense-mediated mRNA decay pathway.
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Upf1p, Nmd2p, and Upf3p are interacting components of the yeast nonsense-mediated mRNA decay pathway.

机译：Upf1p，Nmd2p和Upf3p是酵母无义介导的mRNA衰变途径的相互作用组分。

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Rapid turnover of nonsense-containing mRNAs in Saccharomyces cerevisiae is dependent on Upf1p, Nmd2p, and Upf3p, the products of the UPF1, NMD2/UPF2, and UPF3 genes, respectively. We showed previously that Upf1p and Nmd2p interact and that this interaction is required for nonsense-mediated mRNA decay (F. He and A. Jacobson, Genes Dev. 9:437-454, 1995; F. He, A. H. Brown, and A. Jacobson, RNA 2:153-170, 1996). In this study we have used the yeast two-hybrid system to define other protein-protein interactions among the essential components of this decay pathway. Nmd2p-Upf3p and Upf1p-Upf3p interactions were identified, and the respective domains involved in these interactions were delineated by deletion analysis. The domains of Upf1p and Upf3p putatively involved in their mutual interaction were found to correspond to the domains on the two proteins which interact with Nmd2p, suggesting that Nmd2p bridges Upf1p and Upf3p. This conclusion was reinforced by experiments showing that: (i) deletion of NMD2 completely abolishes interactions between Upf1p and Upf3p and (ii) overexpression of full-length Nmd2p or Nmd2p fragments that retain Upf1p- and Upf3p-interacting domains promotes 10- to 200-fold enhancement of Upf1p-Nmd2p-Upf3p complex formation. These results; the observation that cells harboring either single or multiple deletions of UPF1, NMD2, and UPF3 inhibit nonsense-mediated mRNA decay to the same extent; and an analysis of the possible targets of a dominant-negative NMD2 allele indicate that Upf1p, Nmd2p, Upf3p, and at least one other factor are functionally dependent, interacting components of the yeast nonsense-mediated mRNA decay pathway.
机译：酿酒酵母中的无意义的mRNA的快速更新分别取决于UPf1，NMD2 / UPF2和UPF3基因的产物Upf1p，Nmd2p和Upf3p。先前我们证明了Upf1p和Nmd2p相互作用，并且这种相互作用是无义介导的mRNA衰变所必需的（F.He和A.Jacobson，Genes Dev.9：437-454，1995; F.He，AH Brown和A. Jacobson，RNA 2：153-170，1996）。在这项研究中，我们已经使用酵母双杂交系统定义了该衰变途径的基本组成部分之间的其他蛋白质-蛋白质相互作用。鉴定了Nmd2p-Upf3p和Upf1p-Upf3p相互作用，并通过缺失分析描绘了参与这些相互作用的各个域。发现可能参与相互作用的Upf1p和Upf3p结构域与与Nmd2p相互作用的两种蛋白质上的结构域相对应，这表明Nmd2p桥接Upf1p和Upf3p。实验表明，这一结论进一步证明了这一点：（i）删除NMD2完全消除了Upf1p和Upf3p之间的相互作用，以及（ii）保留了Upf1p和Upf3p相互作用域的全长Nmd2p或Nmd2p片段的过表达促进了10-至200-倍增强Upf1p-Nmd2p-Upf3p复合物的形成。这些结果;观察到带有UPF1，NMD2和UPF3的单个或多个缺失的细胞在相同程度上抑制了无义介导的mRNA衰减;对显性阴性NMD2等位基因可能的靶标的分析表明，Upf1p，Nmd2p，Upf3p和至少一种其他因子是功能依赖性的酵母无义介导的mRNA衰变途径的相互作用组分。

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