Mapping translation 'hot-spots' in live cells by tracking single molecules of mRNA and ribosomes
- Albert Einstein College of Medicine, United States
- Janelia Research Campus, Howard Hughes Medical Institute, United States
- Stanford University School of Medicine, United States
- Massachusetts Institute of Technology, United States
Abstract
Messenger RNA localization is important for cell motility by local protein translation. However, while single mRNAs can be imaged and their movements tracked in single cells, it has not yet been possible to determine whether these mRNAs are actively translating. Therefore, we imaged single β-actin mRNAs tagged with MS2 stem loops colocalizing with labeled ribosomes to determine when polysomes formed. A dataset of tracking information consisting of thousands of trajectories per cell demonstrated that mRNAs co-moving with ribosomes have significantly different diffusion properties from non-translating mRNAs that were exposed to translation inhibitors. This data indicates that ribosome load changes mRNA movement and therefore highly translating mRNAs move slower. Importantly, β-actin mRNA near focal adhesions exhibited sub-diffusive corralled movement characteristic of increased translation. This method can identify where ribosomes become engaged for local protein production and how spatial regulation of mRNA-protein interactions mediates cell directionality.
Article and author information
Author details
Copyright
© 2016, Katz et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
Metrics
-
- 10,341
- views
-
- 2,079
- downloads
-
- 115
- citations
Views, downloads and citations are aggregated across all versions of this paper published by eLife.
Download links
A two-part list of links to download the article, or parts of the article, in various formats.
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
Mapping translation 'hot-spots' in live cells by tracking single molecules of mRNA and ribosomes
eLife 5:e10415.
https://doi.org/10.7554/eLife.10415
