Mapping translation 'hot-spots' in live cells by tracking single molecules of mRNA and ribosomes
Name
Katz-2016-Mapping translation.pdf
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5.98 MB
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Adobe PDF
Checksum (MD5)
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Author(s)
Katz, Zachary B
English, Brian P
Lionnet, Timothee
Yoon, Young J
Monnier, Nilah
Ovryn, Ben
Bathe, Mark
Singer, Robert H
Date Issued
January 2016
Journal
eLife
Publisher
eLife Sciences Publications, Ltd.
Citation
Katz, Zachary B, Brian P English, Timothee Lionnet, Young J Yoon, Nilah Monnier, Ben Ovryn, Mark Bathe, and Robert H Singer. “Mapping Translation ‘Hot-Spots’ in Live Cells by Tracking Single Molecules of mRNA and Ribosomes.” eLife 5 (January 13, 2016).
Version
Final published version
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. These data indicate 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.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Terms of Use
Creative Commons Attribution
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.7554/eLife.10415
