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Structural dynamics of ribosome subunit association studied by mixing-spraying time-resolved cryogenic electron microscopy.

ABSTRACT: Ribosomal subunit association is a key checkpoint in translation initiation but its structural dynamics are poorly understood. Here, we used a recently developed mixing-spraying, time-resolved, cryogenic electron microscopy (cryo-EM) method to study ribosomal subunit association in the sub-second time range. We have improved this method and increased the cryo-EM data yield by tenfold. Pre-equilibrium states of the association reaction were captured by reacting the mixture of ribosomal subunits for 60 ms and 140 ms. We also identified three distinct ribosome conformations in the associated ribosomes. The observed proportions of these conformations are the same in these two time points, suggesting that ribosomes equilibrate among the three conformations within less than 60 ms upon formation. Our results demonstrate that the mixing-spraying method can capture multiple states of macromolecules during a sub-second reaction. Other fast processes, such as translation initiation, decoding, and ribosome recycling, are amenable to study with this method.

SUBMITTER: Chen B

PROVIDER: S-EPMC4456197 | biostudies-literature | 2015 Jun

REPOSITORIES: biostudies-literature

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Structural dynamics of ribosome subunit association studied by mixing-spraying time-resolved cryogenic electron microscopy.

Chen Bo B Kaledhonkar Sandip S Sun Ming M Shen Bingxin B Lu Zonghuan Z Barnard David D Lu Toh-Ming TM Gonzalez Ruben L RL Frank Joachim J

Structure (London, England : 1993) 20150521 6

Ribosomal subunit association is a key checkpoint in translation initiation but its structural dynamics are poorly understood. Here, we used a recently developed mixing-spraying, time-resolved, cryogenic electron microscopy (cryo-EM) method to study ribosomal subunit association in the sub-second time range. We have improved this method and increased the cryo-EM data yield by tenfold. Pre-equilibrium states of the association reaction were captured by reacting the mixture of ribosomal subunits f ...[more]

PMID: 26004440

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Dataset Information

Structural dynamics of ribosome subunit association studied by mixing-spraying time-resolved cryogenic electron microscopy.

Publications

Structural dynamics of ribosome subunit association studied by mixing-spraying time-resolved cryogenic electron microscopy.

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