Journal
JOURNAL OF CELL BIOLOGY
Volume 214, Issue 6, Pages 769-781Publisher
ROCKEFELLER UNIV PRESS
DOI: 10.1083/jcb.201605024
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Funding
- Ministere de l'Education Nationale de la Recherche et de Technologie fellowship
- Site de Recherche Integree sur le Cancer (SIRIC) Montpellier Cancer [INCa-DGOS-INSERM 6045]
- Fondation de la Recherche Medicale grant
- Agence Nationale de la Recherche HI-FISH grant
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Translation is an essential step in gene expression. In this study, we used an improved SunTag system to label nascent proteins and image translation of single messenger ribonucleoproteins (mRNPs) in human cells. Using a dedicated reporter RNA, we observe that translation of single mRNPs stochastically turns on and off while they diffuse through the cytoplasm. We further measure a ribosome density of 1.3 per kilobase and an elongation rate of 13-18 amino acids per second. Tagging the endogenous POLR2A gene revealed similar elongation rates and ribosomal densities and that nearly all messenger RNAs (mRNAs) are engaged in translation. Remarkably, tagging of the heavy chain of dynein 1 (DYNC1H1) shows this mRNA accumulates in foci containing three to seven RNA molecules. These foci are translation sites and thus represent specialized translation factories. We also observe that DYNC1H1 polysomes are actively transported by motors, which may deliver the mature protein at appropriate cellular locations. The SunTag should be broadly applicable to study translational regulation in live single cells.
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