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古菌中的硒蛋白合成:詹氏甲烷球菌中一个可能指导硒代半胱氨酸插入的mRNA元件的鉴定。

Selenoprotein synthesis in archaea: identification of an mRNA element of Methanococcus jannaschii probably directing selenocysteine insertion.

作者信息

Wilting R, Schorling S, Persson B C, Böck A

机构信息

Lehrstuhl für Mikrobiologie der Universität München, Germany.

出版信息

J Mol Biol. 1997 Mar 7;266(4):637-41. doi: 10.1006/jmbi.1996.0812.

Abstract

Selenocysteine is encoded by a UGA codon in all organisms that synthesise selenoproteins. This codon is specified as a selenocysteine codon by an mRNA secondary structure, which is located immediately 3' of the UGA in the reading frame of selenoprotein genes in Gram-negative bacteria, whereas it is located in the 3' untranslated region of eukaryal selenoprotein genes. The location and the structure of a similar mRNA signal in archaea has so far not been determined. Seven selenoproteins were identified for the archaeon Methanococcus jannaschii by labelling with 75Se and by SDS/polyacrylamide electrophoresis. Their size could be correlated with open reading frames possessing internal UGA codons from the total genomic sequence. One of the open reading frames, that of the VhuD subunit of a hydrogenase, possesses two UGA codons and appears to code for a selenoprotein with two selenocysteine residues. A strongly conserved mRNA element was identified that is exclusively linked to selenoprotein genes. It is located in the 3' untranslated region in six of the mRNAs and in the 5' untranslated region of the fdhA mRNA. This element, which is present in the 3' non-translated region of two selenoprotein mRNAs from Methanococcus voltae, is proposed to act in decoding of the UGA with selenocysteine.

摘要

在所有合成硒蛋白的生物体中,硒代半胱氨酸由UGA密码子编码。在革兰氏阴性细菌中,该密码子在硒蛋白基因阅读框中UGA的紧接3'端的mRNA二级结构作用下被指定为硒代半胱氨酸密码子,而在真核生物硒蛋白基因中,它位于3'非翻译区。古菌中类似mRNA信号的位置和结构迄今尚未确定。通过用75Se标记和SDS/聚丙烯酰胺电泳,为詹氏甲烷球菌鉴定出了7种硒蛋白。它们的大小与来自全基因组序列的含有内部UGA密码子的开放阅读框相关。其中一个开放阅读框,即氢化酶VhuD亚基的开放阅读框,含有两个UGA密码子,似乎编码一种含有两个硒代半胱氨酸残基的硒蛋白。鉴定出了一个与硒蛋白基因专门相关的高度保守的mRNA元件。它位于6种mRNA的3'非翻译区和fdhA mRNA的5'非翻译区。存在于沃氏甲烷球菌两种硒蛋白mRNA的3'非翻译区的这个元件,被认为在UGA密码子解码为硒代半胱氨酸的过程中发挥作用。

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