@article {68462,
	title = {Cap-dependent translation initiation monitored in living cells.},
	journal = {Nature Communications},
	volume = {13},
	year = {2022},
	month = {2022 Nov 02},
	pages = {6558},
	abstract = {<p>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.</p>
},
	keywords = {Peptide Chain Initiation, Translational, Peptide Initiation Factors, Protein Biosynthesis, RNA Caps, RNA, Messenger},
	issn = {2041-1723},
	doi = {10.1038/s41467-022-34052-8},
	author = {Gandin, Valentina and English, Brian P and Freeman, Melanie and Leroux, Louis-Philippe and Preibisch, Stephan and Walpita, Deepika and Jaramillo, Maritza and Singer, Robert H}
}
@article {65281,
	title = {Correlative three-dimensional super-resolution and block-face electron microscopy of whole vitreously frozen cells.},
	journal = {Science},
	volume = {367},
	year = {2020},
	month = {2020 Jan 17},
	abstract = {<p>Within cells, the spatial compartmentalization of thousands of distinct proteins serves a multitude of diverse biochemical needs. Correlative super-resolution (SR) fluorescence and electron microscopy (EM) can elucidate protein spatial relationships to global ultrastructure, but has suffered from tradeoffs of structure preservation, fluorescence retention, resolution, and field of view. We developed a platform for three-dimensional cryogenic SR and focused ion beam-milled block-face EM across entire vitreously frozen cells. The approach preserves ultrastructure while enabling independent SR and EM workflow optimization. We discovered unexpected protein-ultrastructure relationships in mammalian cells including intranuclear vesicles containing endoplasmic reticulum-associated proteins, web-like adhesions between cultured neurons, and chromatin domains subclassified on the basis of transcriptional activity. Our findings illustrate the value of a comprehensive multimodal view of ultrastructural variability across whole cells.</p>
},
	issn = {1095-9203},
	doi = {10.1126/science.aaz5357},
	author = {Hoffman, David P and Shtengel, Gleb and Xu, C Shan and Campbell, Kirby R and Freeman, Melanie and Wang, Lei and Milkie, Daniel E and Pasolli, H Amalia and Iyer, Nirmala and Bogovic, John A and Stabley, Daniel R and Shirinifard, Abbas and Pang, Song and Peale, David and Schaefer, Kathy and Pomp, Wim and Chang, Chi-Lun and Lippincott-Schwartz, Jennifer and Kirchhausen, Tom and Solecki, David J and Betzig, Eric and Hess, Harald F}
}
