Ensemble and time-resolved cryo-EM in transcription and translation
| Authors | |
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| Year of publication | 2022 |
| Type | Conference abstract |
| MU Faculty or unit | |
| Citation | |
| Description | During protein synthesis, transfer RNAs (tRNAs) and messenger RNA (mRNA) codons are delivered and translocated within the ribosome from the A to P to E sites, respectively. The tRNA delivery, tRNA proof-reading and translocation of the tRNA anticodons and mRNA are catalyzed by conserved GTPases, elongation factor Tu and G (EF-Tu, EF-G) in bacteria. The structural mechanisms how the ribosome and elongation factors maintain the open reading frame has not been visualized because the rapid GTP hydrolysis step has prevented the capture of authentic EF-G-bound structural intermediates. The lecture will be aimed on the usage of time-resolved cryo-EM to characterize tRNA delivery/proof-reading and translocation in time dependent manner. We will report newly described intermediate structural states, which visualized the delivery and transition of the tRNAs from the A and P to P and E sites during GTP hydrolysis on EF-Tu and EF-G. Moreover, we will highlight how time-resolved cryo-EM can be utilized towards the mechanistic characterization of more complex biological assemblies like coupled transcription-translation in bacteria. |
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