Gu H, Das Gupta J, Schoenberg D R
Department of Pharmacology, Ohio State Biochemistry Program, and the Comprehensive Cancer Center, The Ohio State University, Columbus, OH 43210, USA.
Proc Natl Acad Sci U S A. 1999 Aug 3;96(16):8943-8. doi: 10.1073/pnas.96.16.8943.
Most vertebrate mRNAs exit the nucleus with a 200+-residue poly(A) tail and are deadenylated to yield heterogeneous polymers of 50-200 adenosine residues on any given mRNA. We previously reported that Xenopus albumin mRNA and pre-mRNA have an unusually short, discrete 17-residue poly(A) tail and showed that regulation of poly(A) length is controlled independently by two cis-acting poly(A)-limiting elements (PLE A and PLE B) located in the terminal exon. The present study sought to determine the generality of this regulatory mechanism. Transferrin mRNA also has a discrete <20-nt poly(A) tail, and deletion mapping experiments identified an element homologous to the albumin gene PLE B within the terminal exon of the transferrin gene that conferred poly(A) length regulation on a globin reporter mRNA. Based on this similarity the PLE B sequence was used in a database search to identify candidate mRNA targets for regulated polyadenylation. Of the several hundred sequences identified in this manner we focused on HIV-EP2/Schnurri-2, a member of a family of genes encoding related zinc finger transcription factors. A striking feature of the PLE-like element in these genes is its location 10-33 bp upstream of the translation stop codon. We demonstrate that HIV-EP2 mRNA has a <20-nt poly(A) tail, for which the identified PLE-like sequence is responsible. These results indicate that the presence of a PLE can predict mRNAs with <20-nt poly(A) tails, and that nuclear regulation of poly(A) tail length is a feature of many mRNAs.
大多数脊椎动物的信使核糖核酸(mRNA)带着一条200多个残基的聚腺苷酸(poly(A))尾巴离开细胞核,随后被去腺苷酸化,从而在任何给定的mRNA上产生由50 - 200个腺苷残基组成的异质聚合物。我们之前报道过,非洲爪蟾白蛋白mRNA和前体mRNA具有异常短且离散的17个残基的聚(A)尾巴,并表明聚(A)长度的调节由位于末端外显子中的两个顺式作用聚(A)限制元件(PLE A和PLE B)独立控制。本研究旨在确定这种调节机制的普遍性。转铁蛋白mRNA也有一个离散的小于20个核苷酸的聚(A)尾巴,缺失定位实验在转铁蛋白基因的末端外显子中鉴定出一个与白蛋白基因PLE B同源的元件,该元件赋予了对球蛋白报告mRNA的聚(A)长度调节作用。基于这种相似性,PLE B序列被用于数据库搜索,以识别受调控聚腺苷酸化的候选mRNA靶点。以这种方式鉴定出的数百个序列中,我们重点关注了HIV-EP2/Schnurri-2,它是一个编码相关锌指转录因子的基因家族的成员。这些基因中类似PLE的元件的一个显著特征是其位于翻译终止密码子上游10 - 33个碱基对处。我们证明HIV-EP2 mRNA有一个小于20个核苷酸的聚(A)尾巴,所鉴定的类似PLE的序列对此负责。这些结果表明,PLE的存在可以预测具有小于20个核苷酸聚(A)尾巴的mRNA,并且聚(A)尾巴长度的核调节是许多mRNA的一个特征。