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

Chong Wang; Boran Han; Ruobo Zhou; Xiaowei Zhuang

doi:10.1016/j.cell.2016.04.040

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

Cell. 2016 May 5;165(4):990-1001. doi: 10.1016/j.cell.2016.04.040.

Authors

Chong Wang¹, Boran Han¹, Ruobo Zhou¹, Xiaowei Zhuang²

Affiliations

¹ Howard Hughes Medical Institute, Department of Chemistry and Chemical Biology and Department of Physics, Harvard University, Cambridge, MA 02138, USA.
² Howard Hughes Medical Institute, Department of Chemistry and Chemical Biology and Department of Physics, Harvard University, Cambridge, MA 02138, USA. Electronic address: zhuang@chemistry.harvard.edu.

Abstract

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Dendrites / metabolism
Humans
Optical Imaging / methods*
Polyribosomes / metabolism
Protein Biosynthesis* / drug effects
Protein Synthesis Inhibitors / pharmacology
RNA, Messenger / chemistry
RNA, Messenger / metabolism*

Substances

Protein Synthesis Inhibitors
RNA, Messenger

Grants and funding

R01 GM096450/GM/NIGMS NIH HHS/United States