Sequences mediating the translation of mouse S16 ribosomal protein mRNA during myoblast differentiation and in vitro and possible control points for the in vitro translation.
Sequences mediating the translation of mouse S16 ribosomal protein mRNA during myoblast differentiation and in vitro and possible control points for the in vitro translation.
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成肌细胞分化和体外介导小鼠 S16 核糖体蛋白 mRNA 翻译的序列以及体外翻译的可能控制点。
DOI：
10.1101/gad.5.9.1723
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发表时间：
1991
影响因子：
10.5
通讯作者：
Bowman,LH
中科院分区：
文献类型：
--
作者：
Hammond,ML;Merrick,W;Bowman,LH
核糖体蛋白（r-蛋白）mRNA的翻译通常是低效的，并在小鼠成肌细胞分化为纤维的过程中受到调节。在本讨论中，我们表明，S16 r蛋白mRNA的前31个核苷酸，当位于mRNA的5'端时，足以将r蛋白mRNA的翻译特性赋予SV-GALK mRNA，其通常在成肌细胞和纤维中有效翻译。然而，如果相同的S16序列位于SV-GALK mRNA的5 ′-非翻译区的内部，则它们不赋予SV-GALK mRNA r-蛋白mRNA的翻译性质。在体外检查小鼠r蛋白mRNA的翻译，以帮助阐明控制其翻译的机制。小鼠r蛋白mRNA在兔网织红细胞提取物中翻译效率低下，在成肌细胞分化过程中介导其低效和受调控翻译的相同序列也以位置依赖性方式介导其在网织红细胞提取物中的低效翻译。为了确定在体内不主动翻译的亚多核糖体r蛋白mRNA是否能够翻译，在网织红细胞提取物中翻译亚多核糖体RNA。亚多核糖体r蛋白mRNA与多核糖体mRNA一样具有翻译能力。为了帮助鉴定起始因子和/或起始途径中介导r蛋白mRNA低效翻译的步骤，网织红细胞提取物补充了纯化的起始因子。只有eIF-4F（帽结合复合物）和eIF-3（参与亚基解离并在起始过程中与eIF-4F相互作用）刺激r蛋白mRNA的翻译。这些实验，沿着m7 GDP抑制研究，表明eIF-4 F和/或eIF-3，或由这些因子介导的步骤，介导网织红细胞提取物中的低效翻译，并提高了这些步骤也控制成肌细胞分化期间r蛋白mRNA的调节翻译的可能性。
The translation of ribosomal protein (r-protein) mRNAs is generally inefficient and regulated during the differentiation of mouse myoblasts into fibers. In this discussion we show that the first 31 nucleotides of the S16 r-protein mRNA, when located at the 5' end of the mRNA, are sufficient to impart the translational properties of an r-protein mRNA to the SV-GALK mRNA, which is normally translated efficiently in both myoblasts and fibers. If the same S16 sequences are located within the interior of the 5'-untranslated region of the SV-GALK mRNA, however, they do not impart the translational properties of an r-protein mRNA to the SV-GALK mRNA. The translation of mouse r-protein mRNAs was examined in vitro to help elucidate the mechanisms controlling their translation. Mouse r-protein mRNAs are inefficiently translated in rabbit reticulocyte extracts, and the same sequences that mediate their inefficient and regulated translation during myoblast differentiation also mediate their inefficient translation in a position-dependent manner in reticulocyte extracts. To determine whether the subpolysomal r-protein mRNAs that are not actively translated in vivo are capable of translation, subpolysomal RNA was translated in reticulocyte extracts. The subpolysomal r-protein mRNAs are just as capable of translation as are polysomal mRNAs. To help identify the initiation factors and/or the steps in the initiation pathway that mediate the inefficient translation of r-protein mRNAs, reticulocyte extracts were supplemented with purified initiation factors. Only eIF-4F, the cap-binding complex, and eIF-3, which is involved in subunit dissociation and interacts with eIF-4F during initiation, stimulated the translation of r-protein mRNA. These experiments, along with m7GDP inhibition studies, suggest that eIF-4F and/or eIF-3, or the steps mediated by these factors, mediate the inefficient translation in reticulocyte extracts and raise the possibility that these steps also control the regulated translation of r-protein mRNAs during myoblast differentiation.
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DOI：
--
发表时间：
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期刊：
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