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首页> 外文期刊>Molecular and Cellular Biology >PCF11 encodes a third protein component of yeast cleavage and polyadenylation factor I.
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PCF11 encodes a third protein component of yeast cleavage and polyadenylation factor I.

机译：PCF11编码酵母裂解和聚腺苷酸化因子I的第三种蛋白质成分。

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Cleavage and polyadenylation factor I (CF I) is one of four factors required in vitro for yeast pre-mRNA 3'-end processing. Two protein components of this factor, encoded by genes RNA14 and RNA15, have already been identified. We describe here another gene, PCF11 (for protein 1 of CF I), that genetically interacts with RNA14 and RNA15 and which presumably codes for a third protein component of CF I. This gene was isolated in a two-hybrid screening designed to identify proteins interacting with Rna14 and Rna15. PCF11 is an essential gene encoding for a protein of 626 amino acids having an apparent molecular mass of 70 kDa. Thermosensitive mutations in PCF11 are synergistically lethal with thermosensitive alleles of RNA14 and RNA15. The Pcf11-2 thermosensitive strain shows a shortening of the poly(A) tails and a strong decrease in the steady-state level of actin transcripts after a shift to the nonpermissive temperature as do the thermosensitive alleles of RNA14 and RNA15. Extracts from the pcf11-1 and pcf11-2 thermosensitive strains and the wild-type strain, when Pcf11 is neutralized by specific antibodies, are deficient in cleavage and polyadenylation. Moreover, fractions obtained by anion-exchange chromatography of extracts from the wild-type strain contain both Pcf11 and Rna15 in the same fractions, as shown by immunoblotting with a Pcf11-specific antibody.
机译：裂解和聚腺苷酸化因子I（CF I）是体外酵母前mRNA 3'末端加工所需的四个因子之一。已经鉴定出该因子的两个蛋白质成分，由基因RNA14和RNA15编码。我们在这里描述了另一个基因PCF11（用于CF I的蛋白1），该基因与RNA14和RNA15发生了遗传相互作用，可能编码了CF I的第三种蛋白成分。该基因是在旨在鉴定蛋白的两杂交筛选中分离的与Rna14和Rna15进行交互。 PCF11是编码表观分子量为70 kDa的626个氨基酸的蛋白质的必需基因。 PCF11中的热敏突变与RNA14和RNA15的热敏等位基因协同致死。 Pcf11-2热敏菌株显示出poly（A）尾巴的缩短，并且在转移到非许可温度后，肌动蛋白转录本的稳态水平大大降低，RNA14和RNA15的热敏等位基因也是如此。 pcf11-1和pcf11-2热敏菌株以及野生型菌株的提取物（当Pcf11被特异性抗体中和时）缺乏裂解和聚腺苷酸化作用。而且，通过用野生型菌株的提取物进行阴离子交换层析得到的级分在相同的级分中同时包含Pcf11和Rna15，如用Pcf11特异性抗体免疫印迹所显示的。

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