Endoh Tamaki, Kawasaki Yu, Sugimoto Naoki
Frontier Institute for Biomolecular Engineering Research (FIBER), Konan University, Kobe, Japan.
Methods. 2013 Nov;64(1):73-8. doi: 10.1016/j.ymeth.2013.05.026. Epub 2013 Jun 7.
mRNA forms various secondary and tertiary structures that affect gene expression. Although structures formed in the untranslated regions (UTRs) of mRNAs that inhibit translation have been characterized, stable mRNA structures in open reading frames (ORFs) may also cause translational halt or slow translation elongation. We previously established a method, termed a synchronized translation assay, that enables time course analysis of single turnover translation elongation. In this method, translation initiation, which is a rate determining step of the translation procedure, can be ignored because all ribosomes are synchronized on a specific position of mRNA before translation elongation is restarted from this position. In this paper, we used the synchronized translation assay to evaluate the effects of a G-quadruplex structure located at various positions within the mRNA ORF on translational halt.
信使核糖核酸（mRNA）形成各种影响基因表达的二级和三级结构。虽然已对信使核糖核酸非翻译区（UTR）中形成的抑制翻译的结构进行了表征，但开放阅读框（ORF）中的稳定信使核糖核酸结构也可能导致翻译停滞或减缓翻译延伸。我们之前建立了一种称为同步翻译分析的方法，该方法能够对单次周转翻译延伸进行时间进程分析。在这种方法中，由于在翻译延伸从该位置重新开始之前，所有核糖体都在信使核糖核酸的特定位置同步，因此可以忽略作为翻译过程速率决定步骤的翻译起始。在本文中，我们使用同步翻译分析来评估位于信使核糖核酸开放阅读框内不同位置的G-四链体结构对翻译停滞的影响。
