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首页> 外文期刊>EMBO Journal >Recognition of polyadenylation sites in yeast pre-mRNAs by cleavage and polyadenylation factor.
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Recognition of polyadenylation sites in yeast pre-mRNAs by cleavage and polyadenylation factor.

机译：识别聚腺苷酸化网站的酵母pre-mRNAs乳沟和聚腺苷酸化因子。

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摘要

Recognition of poly(A) sites in yeast pre-mRNAs is poorly understood. Employing an in vitro cleavage system with cleavage and polyadenylation factor (CPF) and cleavage factor IA we show that the efficiency and positioning elements are dispensable for poly(A)-site recognition within a short CYC1 substrate in vitro. Instead, U-rich elements immediately upstream and downstream of the poly(A) site mediate cleavage-site recognition within CYC1 and ADH1 pre-mRNAs. These elements act in concert with the poly(A) site to produce multiple recognition sites for the processing machinery, since combinations of mutations within these elements were most effective in cleavage inhibition. Intriguingly, introduction of a U-rich element downstream of the GAL7 poly(A) site strongly enhanced cleavage, underscoring the importance of downstream sequences in general. RNA- binding analyses demonstrate that cleavage depends on the recognition of the poly(A)-site region by CPF. Consistent with in vitro results, mutation of sequences upstream and downstream of the poly(A) site affected 3'-end formation in vivo. A model for yeast pre-mRNA cleavage-site recognition outlines an unanticipated high conservation of yeast and mammalian 3'-end processing mechanisms.
机译：承认在酵母pre-mRNAs保利(A)网站知之甚少。系统与解理和聚腺苷酸化因子(CPF)和IA我们表明,劈理因素效率和定位元素可有可无的内聚(A)网站识别短CYC1底物在体外。元素立即上游和下游保利(A)网站调解裂解位点识别在CYC1 ADH1 pre-mRNAs。元素一致行动保利(A)的网站产生多个网站的认可加工机械,因为的组合在这些元素最突变有效地抑制卵裂。引入U-rich元素的下游GAL7保利(A)网站强烈增强的乳沟,强调了下游的重要性序列。证明乳沟取决于保利(一)站点区域的识别论坛。与体外结果一致,变异的保利(A)的序列上游和下游网站3 '端体内形成的影响。对酵母pre-mRNA裂解位点识别概述了一个未预料到的高保护酵母和哺乳动物3 '端处理机制。

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