Hector, R. D. , Burlacu, E., Aitken, S., Le Bihan, T., Tuijtel, M., Zaplatina, A., Cook, A. G. and Granneman, S. (2014) Snapshots of pre-rRNA structural flexibility reveal eukaryotic 40S assembly dynamics at nucleotide resolution. Nucleic Acids Research, 42(19), pp. 12138-12154. (doi: 10.1093/nar/gku815)
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Publisher's URL: http://dx.doi.org/10.1093/nar/gku815
Abstract
Ribosome assembly in eukaryotes involves the activity of hundreds of assembly factors that direct the hierarchical assembly of ribosomal proteins and numerous ribosomal RNA folding steps. However, detailed insights into the function of assembly factors and ribosomal RNA folding events are lacking. To address this, we have developed ChemModSeq, a method that combines structure probing, high-throughput sequencing and statistical modeling, to quantitatively measure RNA structural rearrangements during the assembly of macromolecular complexes. By applying ChemModSeq to purified 40S assembly intermediates we obtained nucleotide-resolution maps of ribosomal RNA flexibility revealing structurally distinct assembly intermediates and mechanistic insights into assembly dynamics not readily observed in cryo-electron microscopy reconstructions. We show that RNA restructuring events coincide with the release of assembly factors and predict that completion of the head domain is required before the Rio1 kinase enters the assembly pathway. Collectively, our results suggest that 40S assembly factors regulate the timely incorporation of ribosomal proteins by delaying specific folding steps in the 3′ major domain of the 20S pre-ribosomal RNA.
Item Type: | Articles |
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Status: | Published |
Refereed: | Yes |
Glasgow Author(s) Enlighten ID: | Hector, Dr Ralph |
Authors: | Hector, R. D., Burlacu, E., Aitken, S., Le Bihan, T., Tuijtel, M., Zaplatina, A., Cook, A. G., and Granneman, S. |
College/School: | College of Medical Veterinary and Life Sciences > School of Psychology & Neuroscience |
Journal Name: | Nucleic Acids Research |
Publisher: | Oxford University Press |
ISSN: | 0305-1048 |
ISSN (Online): | 1362-4962 |
Copyright Holders: | Copyright © 2014 The Authors |
First Published: | First published in Nucleic Acids Research 42(19):12138-12154 |
Publisher Policy: | Reproduced under a Creative Commons License |
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