Real-Time Imaging of Translation on Single mRNA Transcripts in Live Cells

• Published in: Cell Vol. 165; no. 4; pp. 990 - 1001
• Main Authors: Wang, Chong, Han, Boran, Zhou, Ruobo, Zhuang, Xiaowei
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 05.05.2016
• Subjects: active transport; Animals; dendrites; Dendrites - metabolism; fluorescence; Humans; image analysis; messenger RNA; Optical Imaging - methods; polypeptides; polyribosomes; Polyribosomes - metabolism; Protein Biosynthesis - drug effects; Protein Synthesis Inhibitors - pharmacology; RNA, Messenger - chemistry; RNA, Messenger - metabolism; translation (genetics)
• ISSN: 0092-8674; 1097-4172; 1097-4172
• DOI: 10.1016/j.cell.2016.04.040

Translation is under tight spatial and temporal controls to ensure protein production in the right time and place in cells. Methods that allow real-time, high-resolution visualization of translation in live cells are essential for understanding the spatiotemporal dynamics of translation regulation. Based on multivalent fluorescence amplification of the nascent polypeptide signal, we develop a method to image translation on individual mRNA molecules in real time in live cells, allowing direct visualization of translation events at the translation sites. Using this approach, we monitor transient changes of translation dynamics in responses to environmental stresses, capture distinct mobilities of individual polysomes in different subcellular compartments, and detect 3′ UTR-dependent local translation and active transport of polysomes in dendrites of primary neurons. [Display omitted] •Translation activity on individual mRNA molecules is imaged using SunTag•Transient translation responses to environmental stress are captured in real time•Polysomes display distinct mobilities at different subcellular localizations•Local translation and active transport of polysomes are observed in dendrites Real-time imaging of translation activities on individual mRNAs in live cells enables the detection of transient translational responses to environmental stresses, motion dynamics of individual polysomes at different subcellular compartments, and local translation and active transport of polysomes in the dendrites of neurons.