@article{567598e07cf54e0593d4f958a614780d,
title = "Cap-dependent translation initiation monitored in living cells",
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{\textquoteright}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.",
author = "Valentina Gandin and English, {Brian P.} and Melanie Freeman and Leroux, {Louis Philippe} and Stephan Preibisch and Deepika Walpita and Maritza Jaramillo and Singer, {Robert H.}",
note = "Funding Information: This work was funded by the Howard Hughes Medical Institute. M.J. is supported by grants from the Canadian Institute for Health Research (CIHR) and Natural Sciences and Engineering Council (NSERC), and a salary award from the Fonds de Recherche du Qu{\'e}bec en Sant{\'e} (FRQS). We are extremely thankful to all the labs, teams, and shared resources at Janelia for supporting our research: James Zhe Liu and Peng Dong for helping us generate the knock-in mouse Embryonic Stem Cells using CRISPR-Cas9. Kimberly Ritola and Hyun Ah Yi (Viral Tools) for producing lentiviral particles used in this study, Kevin McGowan (Molecular Biology) for analyzing the stability of the ARC mRNA, Kathy Schaefer and Jenny Hagemeier (Cell and Tissue Culture) for their help with single-cell sorting. We are grateful to Jim Cox and Kendra Morris (Vivarium) for mouse colony management. Funding Information: This work was funded by the Howard Hughes Medical Institute. M.J. is supported by grants from the Canadian Institute for Health Research (CIHR) and Natural Sciences and Engineering Council (NSERC), and a salary award from the Fonds de Recherche du Qu{\'e}bec en Sant{\'e} (FRQS). We are extremely thankful to all the labs, teams, and shared resources at Janelia for supporting our research: James Zhe Liu and Peng Dong for helping us generate the knock-in mouse Embryonic Stem Cells using CRISPR-Cas9. Kimberly Ritola and Hyun Ah Yi (Viral Tools) for producing lentiviral particles used in this study, Kevin McGowan (Molecular Biology) for analyzing the stability of the ARC mRNA, Kathy Schaefer and Jenny Hagemeier (Cell and Tissue Culture) for their help with single-cell sorting. We are grateful to Jim Cox and Kendra Morris (Vivarium) for mouse colony management. Publisher Copyright: {\textcopyright} 2022, The Author(s).",
year = "2022",
month = dec,
doi = "10.1038/s41467-022-34052-8",
language = "English (US)",
volume = "13",
journal = "Nature communications",
issn = "2041-1723",
publisher = "Nature Publishing Group",
number = "1",
}