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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory Press
2016
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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. |
format | Online Article Text |
id | pubmed-4803056 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
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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