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Stepwise and dynamic assembly of the earliest precursors of small ribosomal subunits in yeast

The eukaryotic ribosomal RNA (rRNA) is associated cotranscriptionally with numerous factors into an enormous 90S preribosomal particle that conducts early processing of small ribosomal subunits. The assembly pathway and structure of the 90S particle is poorly understood. Here, we affinity-purified a...

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Autores principales: Zhang, Liman, Wu, Chen, Cai, Gaihong, Chen, She, Ye, Keqiong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4803056/
https://www.ncbi.nlm.nih.gov/pubmed/26980190
http://dx.doi.org/10.1101/gad.274688.115
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author Zhang, Liman
Wu, Chen
Cai, Gaihong
Chen, She
Ye, Keqiong
author_facet Zhang, Liman
Wu, Chen
Cai, Gaihong
Chen, She
Ye, Keqiong
author_sort Zhang, Liman
collection PubMed
description The eukaryotic ribosomal RNA (rRNA) is associated cotranscriptionally with numerous factors into an enormous 90S preribosomal particle that conducts early processing of small ribosomal subunits. The assembly pathway and structure of the 90S particle is poorly understood. Here, we affinity-purified and analyzed the constituents of yeast 90S particles that were assembled on a series of plasmid-encoded 3′-truncated pre-18S RNAs. We determined the assembly point of 65 proteins and the U3, U14, and snR30 small nucleolar RNAs (snoRNAs), revealing a stepwise and dynamic assembly map. The 5′ external transcribed spacer (ETS) alone can nucleate a large complex. When the 18S rRNA is nearly complete, the 90S structure undergoes a dramatic reorganization, releasing U14, snR30, and 14 protein factors that bind earlier. We also identified a reference state of 90S that is fully assembled yet has not undergone 5′ETS processing. The assembly map present here provides a new framework to understand small subunit biogenesis.
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spelling pubmed-48030562016-09-15 Stepwise and dynamic assembly of the earliest precursors of small ribosomal subunits in yeast Zhang, Liman Wu, Chen Cai, Gaihong Chen, She Ye, Keqiong Genes Dev Research Paper The eukaryotic ribosomal RNA (rRNA) is associated cotranscriptionally with numerous factors into an enormous 90S preribosomal particle that conducts early processing of small ribosomal subunits. The assembly pathway and structure of the 90S particle is poorly understood. Here, we affinity-purified and analyzed the constituents of yeast 90S particles that were assembled on a series of plasmid-encoded 3′-truncated pre-18S RNAs. We determined the assembly point of 65 proteins and the U3, U14, and snR30 small nucleolar RNAs (snoRNAs), revealing a stepwise and dynamic assembly map. The 5′ external transcribed spacer (ETS) alone can nucleate a large complex. When the 18S rRNA is nearly complete, the 90S structure undergoes a dramatic reorganization, releasing U14, snR30, and 14 protein factors that bind earlier. We also identified a reference state of 90S that is fully assembled yet has not undergone 5′ETS processing. The assembly map present here provides a new framework to understand small subunit biogenesis. Cold Spring Harbor Laboratory Press 2016-03-15 /pmc/articles/PMC4803056/ /pubmed/26980190 http://dx.doi.org/10.1101/gad.274688.115 Text en © 2016 Zhang et al.; Published by Cold Spring Harbor Laboratory Press http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
spellingShingle Research Paper
Zhang, Liman
Wu, Chen
Cai, Gaihong
Chen, She
Ye, Keqiong
Stepwise and dynamic assembly of the earliest precursors of small ribosomal subunits in yeast
title Stepwise and dynamic assembly of the earliest precursors of small ribosomal subunits in yeast
title_full Stepwise and dynamic assembly of the earliest precursors of small ribosomal subunits in yeast
title_fullStr Stepwise and dynamic assembly of the earliest precursors of small ribosomal subunits in yeast
title_full_unstemmed Stepwise and dynamic assembly of the earliest precursors of small ribosomal subunits in yeast
title_short Stepwise and dynamic assembly of the earliest precursors of small ribosomal subunits in yeast
title_sort stepwise and dynamic assembly of the earliest precursors of small ribosomal subunits in yeast
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4803056/
https://www.ncbi.nlm.nih.gov/pubmed/26980190
http://dx.doi.org/10.1101/gad.274688.115
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