ArticleProceedings of the National Academy of Sciences of the United States of America2014
Initial bridges between two ribosomal subunits are formed within 9.4 milliseconds, as studied by time-resolved cryo-EM.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed.
- Article
- Passage of ribosomes through microsprayer increases functional activity - Implications for activity assays in time-resolved cryo-EM.Journal of structural biology · 2025Article
- Frozen in time: analyzing molecular dynamics with time-resolved cryo-EM.Structure (London, England : 1993) · 2023Review
- Exploring the Structural Variability of Dynamic Biological Complexes by Single-Particle Cryo-Electron Microscopy.Micromachines · 2022Review
- Structural dynamics: review of time-resolved cryo-EM.Acta crystallographica. Section D, Structural biology · 2022Review
- Microsecond melting and revitrification of cryo samples: protein structure and beam-induced motion.Acta crystallographica. Section D, Structural biology · 2022Article
- Microsecond melting and revitrification of cryo samples.Structural dynamics (Melville, N.Y.) · 2021Article
- Integrative approaches in cryogenic electron microscopy: Recent advances in structural biology and future perspectives.iScience · 2021Review
- Microscale Fluid Behavior during Cryo-EM Sample Blotting.Biophysical journal · 2020Article
- Joachim Frank's Binding with the Ribosome.Structure (London, England : 1993) · 2019Review
- Biological Applications at the Cutting Edge of Cryo-Electron Microscopy.Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada · 2018Review
- Cryo-EM in drug discovery: achievements, limitations and prospects.Nature reviews. Drug discovery · 2018Review
- How Messenger RNA and Nascent Chain Sequences Regulate Translation Elongation.Annual review of biochemistry · 2018Review
- RNA Structural Dynamics As Captured by Molecular Simulations: A Comprehensive Overview.Chemical reviews · 2018Review
- Never at rest: insights into the conformational dynamics of ion channels from cryo-electron microscopy.The Journal of physiology · 2018Review
- Time-resolved cryo-electron microscopy: Recent progress.Journal of structural biology · 2017Review
- The kinetic mechanism of bacterial ribosome recycling.Nucleic acids research · 2017Article
- A Fast and Effective Microfluidic Spraying-Plunging Method for High-Resolution Single-Particle Cryo-EM.Structure (London, England : 1993) · 2017Article
- Key Intermediates in Ribosome Recycling Visualized by Time-Resolved Cryoelectron Microscopy.Structure (London, England : 1993) · 2016Article
- Whither Ribosome Structure and Dynamics Research? (A Perspective).Journal of molecular biology · 2016Review
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8 authors.
Funding
Abstract
Association of the two ribosomal subunits during the process of translation initiation is a crucial step of protein synthesis. The two subunits (30S and 50S) of the bacterial 70S ribosome are held together by 12 dynamic bridges involving RNA-RNA, RNA-protein, and protein-protein interactions. The process of bridge formation, such as whether all these bridges are formed simultaneously or in a sequential order, is poorly understood. To understand such processes, we have developed and implemented a class of microfluidic devices that mix two components to completion within 0.4 ms and spray the mixture in the form of microdroplets onto an electron microscopy grid, yielding a minimum reaction time of 9.4 ms before cryofixation. Using these devices, we have obtained cryo-EM data corresponding to reaction times of 9.4 and 43 ms and have determined 3D structures of ribosomal subunit association intermediates. Molecular analyses of the cryo-EM maps reveal that eight intersubunit bridges (bridges B1a, B1b, B2a, B2b, B3, B7a, B7b, and B8) form within 9.4 ms, whereas the remaining four bridges (bridges B2c, B4, B5, and B6) take longer than 43 ms to form, suggesting that bridges are formed in a stepwise fashion. Our approach can be used to characterize sequences of various dynamic functional events on complex macromolecular assemblies such as ribosomes.
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