Parallel measurement of dynamic changes in translation rates in single cells.
Parallel measurement of dynamic changes in translation rates in single cells.
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DOI：
10.1038/nmeth.2729
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发表时间：
2014-01
期刊：
影响因子：
48
通讯作者：
Meyer, Tobias
中科院分区：
文献类型：
--
作者：
Han, Kyuho;Jaimovich, Ariel;Dey, Gautam;Ruggero, Davide;Meyuhas, Oded;Sonenberg, Nahum;Meyer, Tobias
蛋白质浓度通常由翻译速率的动态变化来调节。然而，直接监测活细胞中翻译的变化一直是一个挑战。我们开发了一种报告系统，通过将翻译调控基序与编码核靶向荧光蛋白和可控不稳定结构域的序列结合，来测量人类或小鼠单个细胞中翻译速率的实时变化。该方法的应用表明，单个细胞的翻译速率经历了显着的波动，其5‘末端寡核苷酸(5’TOP)基序调节核糖体蛋白的合成。此外，我们发现氨基酸水平的轻微下降通过不同的mTOR依赖的途径来控制top mRNA的翻译，而氨基酸水平的较大下降通过eIF2A控制翻译。我们的研究表明，动态测量翻译调控基元的单细胞活动可以用来识别和研究翻译的基本原则。
Protein concentrations are often regulated by dynamic changes in translation rates. Nevertheless, it has been challenging to directly monitor changes in translation in living cells. We have developed a reporter system to measure real-time changes of translation rates in human or mouse individual cells by conjugating translation regulatory motifs to sequences encoding a nuclear targeted fluorescent protein and a controllable destabilization domain. Application of the method showed that individual cells undergo marked fluctuations in the translation rate of mRNAs whose 5′ terminal oligopyrimidine (5′ TOP) motif regulates the synthesis of ribosomal proteins. Furthermore, we show that small reductions in amino acid levels signal through different mTOR-dependent pathways to control TOP mRNA translation, whereas larger reductions in amino acid levels control translation through eIF2A. Our study demonstrates that dynamic measurements of single-cell activities of translation regulatory motifs can be used to identify and investigate fundamental principles of translation.
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影响因子：
64.5
作者：
Hafner M;Landthaler M;Burger L;Khorshid M;Hausser J;Berninger P;Rothballer A;Ascano M Jr;Jungkamp AC;Munschauer M;Ulrich A;Wardle GS;Dewell S;Zavolan M;Tuschl T
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Tuschl T
影响因子：
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Sonenberg, Nahum
影响因子：
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影响因子：
64.8
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影响因子：
5.3
作者：
GEYER, PK;MEYUHAS, O;JOHNSON, LF
通讯作者：
JOHNSON, LF