RT Journal Article
SR Electronic
T1 Multi-color single molecule imaging uncovers extensive heterogeneity in mRNA decoding
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 477661
DO 10.1101/477661
A1 Boersma, Sanne
A1 Khuperkar, Deepak
A1 Verhagen, Bram M.P.
A1 Sonneveld, Stijn
A1 Grimm, Jonathan B.
A1 Lavis, Luke D.
A1 Tanenbaum, Marvin E.
YR 2018
UL http://biorxiv.org/content/early/2018/11/24/477661.abstract
AB mRNA translation is a key step in decoding genetic information. Genetic decoding is surprisingly heterogeneous, as multiple distinct polypeptides can be synthesized from a single mRNA sequence. To study translational heterogeneity, we developed the MoonTag, a new fluorescence labeling system to visualize translation of single mRNAs. When combined with the orthogonal SunTag system, the MoonTag enables dual readouts of translation, greatly expanding the possibilities to interrogate complex translational heterogeneity. By placing MoonTag and SunTag sequences in different translation reading frames, each driven by distinct translation start sites, start site selection of individual ribosomes can be visualized in real-time. We find that start site selection is largely stochastic, but that the probability of using a particular start site differs among mRNA molecules, and can be dynamically regulated over time. Together, this study provides key insights into translation start site selection heterogeneity, and provides a powerful toolbox to visualize complex translation dynamics.