The 3′ untranslated region of manganese superoxide dismutase RNA contains a translational enhancer element
The 3′ untranslated region of manganese superoxide dismutase RNA contains a translational enhancer element
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DOI：
10.1021/bi980935g
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发表时间：
1998-11-17
期刊：
影响因子：
2.9
通讯作者：
Clerch, LB
中科院分区：
文献类型：
--
作者：
Chung, DJ;Wright, AE;Clerch, LB
一种氧化还原敏感蛋白与锰超氧化物歧化酶(MnSOD)RNA的3‘非翻译区(UTR)结合已被描述[Fazzone，H.，Wangner，A.，and Clerch，L.B.(1993)J.Clin]。投资。92,1278-1281；Chung，D.J.和Clerch，L.B.(1997)AmJ.Physiol.16、L714-L719]。在本研究中，通过交叉竞争凝胶延迟和RNase H分析，确定了位于终止密码子下游111个碱基的41个碱基区为参与蛋白质结合的3‘非编码区顺式元件。该区域的碱基序列与在终止密码子下游大致相同距离的大鼠、小鼠、牛和人的MnSODmRNAs中75%保守的3‘UTRs相似。在体外兔网织红细胞裂解物系统中，评估了该蛋白结合区在RNA翻译中的作用。与含有3‘UTR顺式元件的MnSOD RNA相比，缺失3’UTR元件的MnSOD RNA的翻译减少了60%。在抑制MnSODRNA蛋白结合活性的竞争对手寡核苷酸存在下，含有3‘UTR的MnSODRNA的翻译减少了65%。因此，顺式元件和RNA蛋白结合活性都是更有效地翻译MnSOD所必需的。核糖体分析表明，MnSODRNA结合蛋白参与了翻译起始复合体的形成。当MnSODRNA结合活性被抑制时，起始复合体的形成减少了50%。根据本研究的数据，我们认为MnSOD3‘端非编码区顺式元件可能通过与MnSODRNA结合蛋白的相互作用而发挥翻译增强子的作用。
A redox-sensitive protein that binds to the 3' untranslated region (UTR) of manganese superoxide dismutase (MnSOD) RNA has been described previously [Fazzone, H., Wangner, A., and Clerch, L. B. (1993) J. Clin. Invest. 92, 1278-1281; Chung, D. J., and Clerch, L. B. (1997) Am. J. Physiol. 16, L714-L719]. In the present study, cross-competition gel retardation and RNase H assays were used to identify a 41-base region located 111 bases downstream of the stop codon as the 3' UTR cis element involved in protein binding. The base sequence of this region is similar to 75% conserved among the 3' UTRs of rat, mouse, cow, and human MnSOD mRNAs at approximately the same distance downstream of the stop codon. The role of this protein-binding region in RNA translation was assessed in an in vitro rabbit reticulocyte lysate system. Translation of MnSOD RNA from which the 3' UTR element was deleted decreased 60% compared with translation of MnSOD RNA containing the 3' UTR cis element. In the presence of a specific competitor oligoribonucleotide that inhibits MnSOD RNA protein-binding activity, translation of MnSOD RNA containing the 3' UTR was decreased by 65%. Thus, both the cis element and RNA protein-binding activity were required for more efficient translation of the MnSOD. An analysis of ribosomal profiles suggests the MnSOD RNA-binding protein participates in the formation of the translation initiation complex. When MnSOD RNA-binding activity was inhibited, initiation complex formation was decreased by 50%. From the data obtained in this study, we propose that the 3' UTR cis element of MnSOD through its interaction with MnSOD RNA-binding protein may function as a translational enhancer.