eIF5A promotes translation of polyproline motifs.

eIF5A promotes translation of polyproline motifs.
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DOI:
10.1016/j.molcel.2013.04.021
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发表时间:
2013-07-11
期刊:
影响因子:
16
通讯作者:
Dever, Thomas E.
Dever, Thomas E.
中科院分区:
生物学1区
文献类型:
--
作者:
Gutierrez, Erik;Shin, Byung-Sik;Woolstenhulme, Christopher J.;Kim, Joo-Ran;Saini, Preeti;Buskirk, Allen R.;Dever, Thomas E.

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翻译因子eIF 5A,含有独特的氨基酸羟腐胺赖氨酸,最初显示刺激甲硫氨酰嘌呤霉素合成,一种用于肽键形成的模型测定。最近,eIF 5A被证明可以促进翻译延伸;然而,它在蛋白质合成中的精确要求仍然难以捉摸。在这里,我们使用在酵母和体外重建的翻译试验,以揭示一个特定的要求eIF 5A,以促进连续脯氨酸残基之间的肽键形成。eIF 5A的加入缓解了体外三个连续脯氨酸残基翻译过程中的核糖体停滞,并且eIF 5A功能的丧失损害了体内含聚脯氨酸蛋白的翻译。羟基自由基探测实验将eIF 5A定位在核糖体的E位点附近,其羟腐胺赖氨酸残基邻近P位点tRNA的受体茎。因此,eIF 5A,像它的细菌直系同源物EFP,提出刺激核糖体的肽基转移酶活性,并促进穷人的底物,如脯氨酸的反应性。
Translation factor eIF5A, containing the unique amino acid hypusine, was originally shown to stimulate methionyl-puromycin synthesis, a model assay for peptide bond formation. More recently, eIF5A was shown to promote translation elongation; however, its precise requirement in protein synthesis has remained elusive. Here we use in vivo assays in yeast and in vitro reconstituted translation assays to reveal a specific requirement for eIF5A to promote peptide-bond formation between consecutive proline residues. Addition of eIF5A relieves ribosomal stalling during translation of three consecutive proline residues in vitro, and loss of eIF5A function impairs translation of polyproline-containing proteins in vivo. Hydroxyl radical probing experiments localized eIF5A near the E site of the ribosome with its hypusine residue adjacent to the acceptor stem of the P-site tRNA. Thus, eIF5A, like its bacterial ortholog EFP, is proposed to stimulate the peptidyl-transferase activity of the ribosome and facilitate the reactivity of poor substrates like proline.
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发表时间: 2010-09-24
期刊: Molecular cell
影响因子: 16
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