Fagegaltier D, Hubert N, Yamada K, Mizutani T, Carbon P, Krol A
UPR du CNRS Structure des Macromolécules Biologiques et Mécanismes de Reconnaissance, Institut de Biologie Moléculaire et Cellulaire, 15, Rue René Descartes, 67084 Strasbourg Cedex, France.
EMBO J. 2000 Sep 1;19(17):4796-805. doi: 10.1093/emboj/19.17.4796.
Decoding of UGA selenocysteine codons in eubacteria is mediated by the specialized elongation factor SelB, which conveys the charged tRNA(Sec) to the A site of the ribosome, through binding to the SECIS mRNA hairpin. In an attempt to isolate the eukaryotic homolog of SelB, a database search in this work identified a mouse expressed sequence tag containing the complete cDNA encoding a novel protein of 583 amino acids, which we called mSelB. Several lines of evidence enabled us to establish that mSelB is the bona fide mammalian elongation factor for selenoprotein translation: it binds GTP, recognizes the Sec-tRNA(Sec) in vitro and in vivo, and is required for efficient selenoprotein translation in vivo. In contrast to the eubacterial SelB, the recombinant mSelB alone is unable to bind specifically the eukaryotic SECIS RNA hairpin. However, complementation with HeLa cell extracts led to the formation of a SECIS-dependent complex containing mSelB and at least another factor. Therefore, the role carried out by a single elongation factor in eubacterial selenoprotein translation is devoted to two or more specialized proteins in eukaryotes.
真细菌中UGA硒代半胱氨酸密码子的解码由专门的延伸因子SelB介导,该因子通过与SECIS mRNA发夹结构结合,将携带硒代半胱氨酸的tRNA(Sec)转运至核糖体的A位点。为了分离SelB的真核同源物,本研究通过数据库搜索鉴定出一个小鼠表达序列标签,其包含编码一种583个氨基酸的新型蛋白质的完整cDNA,我们将其命名为mSelB。多项证据使我们能够确定mSelB是硒蛋白翻译真正的哺乳动物延伸因子:它结合GTP,在体外和体内识别Sec-tRNA(Sec),并且是体内高效硒蛋白翻译所必需的。与真细菌的SelB不同,单独的重组mSelB无法特异性结合真核SECIS RNA发夹结构。然而,与HeLa细胞提取物互补导致形成了一个包含mSelB和至少另一种因子的SECIS依赖性复合物。因此,在真细菌硒蛋白翻译中由单一延伸因子执行的功能在真核生物中由两种或更多种专门的蛋白质来承担。