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首页> 外文期刊>Molecular and Cellular Biology >Pbp1p, a Factor Interacting withSaccharomyces cerevisiae Poly(A)-Binding Protein, Regulates Polyadenylation
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Pbp1p, a Factor Interacting withSaccharomyces cerevisiae Poly(A)-Binding Protein, Regulates Polyadenylation

机译：Pbp1p，与酿酒酵母Poly（A）结合蛋白相互作用的因子，调节聚腺苷酸化

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The poly(A) tail of an mRNA is believed to influence the initiation of translation, and the rate at which the poly(A) tail is removed is thought to determine how fast an mRNA is degraded. One key factor associated with this 3′-end structure is the poly(A)-binding protein (Pab1p) encoded by the PAB1 gene inSaccharomyces cerevisiae. In an effort to learn more about the functional role of this protein, we used a two-hybrid screen to determine the factor(s) with which it interacts. We identified five genes encoding factors that specifically interact with the carboxy terminus of Pab1p. Of a total of 44 specific clones identified,PBP1 (for Pab1p-binding protein) was isolated 38 times. Of the putative interacting genes examined, PBP1 promoted the highest level of resistance to 3-aminotriazole (>100 mM) in constructs in which HIS3 was used as a reporter. We determined that a fraction of Pbp1p cosediments with polysomes in sucrose gradients and that its distribution is very similar to that of Pab1p. Disruption ofPBP1 showed that it is not essential for viability but can suppress the lethality associated with a PAB1 deletion. The suppression of pab1Δ by pbp1Δ appears to be different from that mediated by other pab1 suppressors, since disruption of PBP1 does not alter translation rates, affect accumulation of ribosomal subunits, change mRNA poly(A) tail lengths, or result in a defect in mRNA decay. Rather, Pbp1p appears to function in the nucleus to promote proper polyadenylation. In the absence of Pbp1p, 3′ termini of pre-mRNAs are properly cleaved but lack full-length poly(A) tails. These effects suggest that Pbp1p may act to repress the ability of Pab1p to negatively regulate polyadenylation.
机译：据信，mRNA的poly（A）尾部会影响翻译的启动，而poly（A）尾部被去除的速率被认为可以确定mRNA降解的速度。与3'端结构有关的一个关键因素是酿酒酵母中 PAB1 基因编码的poly（A）结合蛋白（Pab1p）。为了更多地了解这种蛋白质的功能作用，我们使用了两个杂交筛选来确定与之相互作用的因子。我们确定了五个基因编码因子与Pab1p的羧基末端特异性相互作用。在总共鉴定出的44个特异性克隆中， PBP1 （针对Pab1p结合蛋白）被分离了38次。在检查的推定相互作用基因中， PBP1 在使用 HIS3 作为报告基因的构建体中对3-氨基三唑（> 100 mM）具有最高的抗性。我们确定了一部分Pbp1p与蔗糖梯度中的多核糖体共沉淀，其分布与Pab1p非常相似。破坏 PBP1 显示其对生存力不是必需的，但可以抑制与 PAB1 缺失相关的致死性。由于 PBP1的破坏， pbp1 Δ对 pab1 Δ的抑制作用似乎与其他 pab1 抑制剂介导的抑制作用不同。 em>不会改变翻译速率，不会影响核糖体亚基的积累，不会改变mRNA poly（A）尾部长度，也不会导致mRNA衰变。相反，Pbp1p似乎在细胞核中起作用，以促进适当的聚腺苷酸化。在没有Pbp1p的情况下，pre-mRNA的3'末端会被正确切割，但缺少全长的poly（A）尾巴。这些效果表明，Pbp1p可能起到抑制Pab1p负调节聚腺苷酸化能力的作用。

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