Design considerations for analyzing protein translation regulation by condensates.

Design considerations for analyzing protein translation regulation by condensates.
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DOI:
10.1261/rna.079002.121
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发表时间:
2022-01
期刊:
RNA (New York, N.Y.)
影响因子:
--
通讯作者:
Gladfelter AS
Gladfelter AS
中科院分区:
其他
文献类型:
--
作者:
Roden CA;Gladfelter AS

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含有RNA的生物分子凝聚物的一个建议作用是翻译调节。在几个特定的背景下,翻译已被证明是由相分离蛋白的存在和促进相分离的条件下调制,可能还有更多的等待发现。确定缩合物依赖性翻译规则的一个有力工具是使用工程化的RNA序列,其可以作为翻译效率的报告基因。这一观点将讨论设计特点,考虑在工程RNA报告,以确定在翻译调控相分离的作用。具体来说,我们将涵盖(i)如何工程RNA序列重演天然蛋白质/RNA相互作用,(ii)的优点和缺点,为常用的报告RNA序列,和(iii)重要的控制实验,以区分结合和缩合依赖的翻译抑制。这篇综述的目的是促进忠实的翻译报告基因的设计和应用,以证明生物分子缩合物在翻译中的生理作用。
One proposed role for biomolecular condensates that contain RNA is translation regulation. In several specific contexts, translation has been shown to be modulated by the presence of a phase-separating protein and under conditions which promote phase separation, and likely many more await discovery. A powerful tool for determining the rules for condensate-dependent translation is the use of engineered RNA sequences, which can serve as reporters for translation efficiency. This Perspective will discuss design features to consider in engineering RNA reporters to determine the role of phase separation in translational regulation. Specifically, we will cover (i) how to engineer RNA sequence to recapitulate native protein/RNA interactions, (ii) the advantages and disadvantages for commonly used reporter RNA sequences, and (iii) important control experiments to distinguish between binding- and condensation-dependent translational repression. The goal of this review is to promote the design and application of faithful translation reporters to demonstrate a physiological role of biomolecular condensates in translation.
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