Halstead James M, Lionnet Timothée, Wilbertz Johannes H, Wippich Frank, Ephrussi Anne, Singer Robert H, Chao Jeffrey A
Friedrich Miescher Institute for Biomedical Research, CH-4058 Basel, Switzerland.
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA. Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, NY 10461, USA. Transcription Imaging Consortium, Howard Hughes Medical Institute Janelia Farm Research Campus, Ashburn, VA 20147, USA.
Science. 2015 Mar 20;347(6228):1367-671. doi: 10.1126/science.aaa3380.
Analysis of single molecules in living cells has provided quantitative insights into the kinetics of fundamental biological processes; however, the dynamics of messenger RNA (mRNA) translation have yet to be addressed. We have developed a fluorescence microscopy technique that reports on the first translation events of individual mRNA molecules. This allowed us to examine the spatiotemporal regulation of translation during normal growth and stress and during Drosophila oocyte development. We have shown that mRNAs are not translated in the nucleus but translate within minutes after export, that sequestration within P-bodies regulates translation, and that oskar mRNA is not translated until it reaches the posterior pole of the oocyte. This methodology provides a framework for studying initiation of protein synthesis on single mRNAs in living cells.
对活细胞中单个分子的分析为基本生物学过程的动力学提供了定量见解；然而，信使核糖核酸（mRNA）翻译的动力学仍有待研究。我们开发了一种荧光显微镜技术，可报告单个mRNA分子的首次翻译事件。这使我们能够研究正常生长、应激期间以及果蝇卵母细胞发育过程中翻译的时空调控。我们已经表明，mRNA不在细胞核中翻译，而是在输出后几分钟内开始翻译，在P小体中的隔离调节翻译，并且oskar mRNA直到到达卵母细胞的后极才开始翻译。这种方法为研究活细胞中单个mRNA上蛋白质合成的起始提供了一个框架。
