1. Cap-dependent translation initiation monitored in living cells.
- Author
-
Gandin, Valentina, English, Brian P., Freeman, Melanie, Leroux, Louis-Philippe, Preibisch, Stephan, Walpita, Deepika, Jaramillo, Maritza, and Singer, Robert H.
- Subjects
RNA regulation ,GENETIC translation ,RIBOSOMES ,SPATIAL resolution ,CELL lines - Abstract
mRNA translation is tightly regulated to preserve cellular homeostasis. Despite extensive biochemical, genetic, and structural studies, a detailed understanding of mRNA translation regulation is lacking. Imaging methodologies able to resolve the binding dynamics of translation factors at single-cell and single-mRNA resolution were necessary to fully elucidate regulation of this paramount process. Here live-cell spectroscopy and single-particle tracking were combined to interrogate the binding dynamics of endogenous initiation factors to the 5'cap. The diffusion of initiation factors (IFs) changed markedly upon their association with mRNA. Quantifying their diffusion characteristics revealed the sequence of IFs assembly and disassembly in cell lines and the clustering of translation in neurons. This approach revealed translation regulation at high spatial and temporal resolution that can be applied to the formation of any endogenous complex that results in a measurable shift in diffusion. mRNA translation is tightly regulated to preserve cellular homeostasis. Here live-cell spectroscopy and single-particle tracking are combined to interrogate the binding dynamics of endogenous initiation factors to mRNAs 5'cap, revealing the sequence of translation initiation factors assembly and disassembly; and the clustering of translation in neurons. [ABSTRACT FROM AUTHOR]
- Published
- 2022
- Full Text
- View/download PDF