ISOLATION OF GENOMIC AND CDNA CLONES ENCODING BOVINE POLY(A)-BINDING-PROTEIN-II
ISOLATION OF GENOMIC AND CDNA CLONES ENCODING BOVINE POLY(A)-BINDING-PROTEIN-II
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DOI：
10.1093/nar/23.20.4034
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发表时间：
1995-10-25
影响因子：
14.9
通讯作者：
WAHLE, E
中科院分区：
文献类型：
--
作者：
NEMETH, A;KRAUSE, S;WAHLE, E
分离了牛 Poly(A) 结合蛋白 II (PAB II) 的 cDNA 克隆。它们的序列预测蛋白质为 32.8 kDa，修正了之前对分子量的估计。该蛋白质包含一个假定的 RNP 型 RNA 结合结构域、一个酸性 N 末端和一个碱性 C 末端结构域。对真实 PAB II 的分析与 cDNA 序列的所有预测都非常一致，只是许多精氨酸残基似乎被翻译后修饰。在大肠杆菌中表达的聚(A)结合蛋白II在聚(A)结合和进行性聚腺苷酸化的重建中具有活性，包括聚(A)尾长控制。cDNA克隆在3'非翻译序列中显示出许多潜在的PAB II结合位点。牛 Poly(A)(+) RNA 含有两个与 PAB II 特异性探针杂交的 mRNA。对基因组克隆的分析显示编码序列中有 6 个内含子。修改后的分子量证明了 PAB II 寡聚物的形成，并重新解释了有关该蛋白质与 Poly(A) 结合的早期数据。
cDNA clones for bovine poly(A) binding protein II (PAB II) were isolated. Their sequence predicts a protein of 32.8 kDa, revising earlier estimates of molecular mass, The protein contains one putative RNA-binding domain of the RNP type, an acidic N-terminal and a basic C-terminal domain, Analyses of authentic PAB II were in good agreement with all predictions from the cDNA sequence except that a number of arginine residues appeared to be post-translationally modified. Poly(A) binding protein Il expressed in Escherichia coli was active in poly(A) binding and reconstitution of processive polyadenylation, including poly(A) tail length control, The cDNA clones showed a number of potential PAB II binding sites in the 3' untranslated sequence. Bovine poly(A)(+) RNA contained two mRNAs hybridizing to a PAB II-specific probe, Analysis of a genomic clone revealed six introns in the coding sequence. The revised molecular mass led to a demonstration of PAB II oligomer formation and a reinterpretation of earlier data concerning the protein's binding to poly(A).