Dynamics of Translation of Single mRNA Molecules In Vivo

Xiaowei Yan, Tim A Hoek, Ronald D Vale, Marvin E Tanenbaum

Research output: Contribution to journal/periodicalArticleScientificpeer-review

Abstract

Regulation of mRNA translation, the process by which ribosomes decode mRNAs into polypeptides, is used to tune cellular protein levels. Currently, methods for observing the complete process of translation from single mRNAs in vivo are unavailable. Here, we report the long-term (>1 hr) imaging of single mRNAs undergoing hundreds of rounds of translation in live cells, enabling quantitative measurements of ribosome initiation, elongation, and stalling. This approach reveals a surprising heterogeneity in the translation of individual mRNAs within the same cell, including rapid and reversible transitions between a translating and non-translating state. Applying this method to the cell-cycle gene Emi1, we find strong overall repression of translation initiation by specific 5' UTR sequences, but individual mRNA molecules in the same cell can exhibit dramatically different translational efficiencies. The ability to observe translation of single mRNA molecules in live cells provides a powerful tool to study translation regulation.

Original languageEnglish
Pages (from-to)976-89
Number of pages14
JournalCell
Volume165
Issue number4
DOIs
Publication statusPublished - 05 May 2016

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