Antisense globin RNA in mouse erythroid tissues: Structure, origin, and possible function
Antisense globin RNA in mouse erythroid tissues: Structure, origin, and possible function
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DOI：
10.1073/pnas.93.6.2476
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发表时间：
1996-03-19
影响因子：
11.1
通讯作者：
Rits, S
中科院分区：
文献类型：
--
作者：
Volloch, V;Schweitzer, B;Rits, S
本文描述的实验的目的是测试小鼠红系组织中是否存在反义球蛋白RNA，如果发现，则表征这些分子。本研究采用了多步骤程序，其中通过与确定的核寡核苷酸连接将分子标签附着到细胞RNA上，连接行为保留了RNA分子的末端，这些末端成为细胞RNA和连接的核寡核苷酸之间的连接，它也明确地保留了身份通过所有后续操作，我们将细胞RNA作为有义或反义分子，使用这种方法，我们在贫血小鼠的红系脾细胞和网织红细胞中鉴定并表征了反义β-珠蛋白RNA，我们在本文中表明，反义珠蛋白RNA与剪接珠蛋白mRNA完全互补，表明模板/转录本关系，它在5'端终止于尿苷酸延伸，反映了poly(A)在正义球蛋白 mRNA 的 3' 末端，根据其 3' 末端的结构，反义球蛋白 RNA 可分为三类：与球蛋白 mRNA 精确对应的全尺寸分子、主要缺少 14 个 3' 末端核苷酸的截短分子、以及含有 17 个额外 3' 末端核苷酸的延伸反义 RNA。 全尺寸反义球蛋白 RNA 在其内部包含两个 14 nt 长的互补序列3'-末端片段对应于球蛋白 mRNA 的 5'-非翻译区，这与主要截断的性质一起表明反义 RNA 可能产生新的有义链球蛋白 mRNA 的机制。
The aim of the experiments described in this paper was to test for the presence of antisense globin RNA in mouse erythroid tissues and, if found, to characterize these molecules, The present study made use of a multistep procedure in which a molecular tag is attached to cellular RNA by ligation with a defined ribooligonucleotide, The act of ligation preserves the termini of RNA molecules, which become the junctions between cellular RNAs and the ligated ribooligonucleotide, It also unambiguously preserves the identity of cellular RNA as a sense or antisense molecule through all subsequent manipulations, Using this approach, we identified and characterized antisense beta-globin RNA in erythroid spleen cells and reticulocytes from anemic mice, We show in this paper that the antisense globin RNA is fully complementary to spliced globin mRNA, indicative of the template/transcript relationship, It terminates at the 5' end with a uridylate stretch, reflecting the presence of poly(A) at the 3' end of the sense globin mRNA, With respect to the structure of their 3' termini, antisense globin RNA can be divided into three categories: full-size molecules corresponding precisely to globin mRNA, truncated molecules lacking predominantly 14 3'-terminal nucleotides, and extended antisense RNA containing 17 additional 3'-terminal nucleotides, The full-size antisense globin RNA contains two 14-nt-long complementary sequences within its 3'-terminal segment corresponding to the 5'-untranslated region of globin mRNA, This, together with the nature of the predominant truncation, suggests a mechanism by which antisense RNA might give rise to new sense-strand globin mRNA.