A pre-ribosomal RNA interaction network involving snoRNAs and the Rok1 helicase

  • Ribosome biogenesis in yeast requires 75 small nucleolar RNAs (snoRNAs) and a myriad of cofactors for processing, modification, and folding of the ribosomal RNAs (rRNAs). For the 19 RNA helicases implicated in ribosome synthesis, their sites of action and molecular functions have largely remained unknown. Here, we have used UV cross-linking and analysis of cDNA (CRAC) to reveal the pre-rRNA binding sites of the RNA helicase Rok1, which is involved in early small subunit biogenesis. Several contact sites were identified in the 18S rRNA sequence, which interestingly all cluster in the “foot” region of the small ribosomal subunit. These include a major binding site in the eukaryotic expansion segment ES6, where Rok1 is required for release of the snR30 snoRNA. Rok1 directly contacts snR30 and other snoRNAs required for pre-rRNA processing. Using cross-linking, ligation and sequencing of hybrids (CLASH) we identified several novel pre-rRNA base-pairing sites for the snoRNAs snR30, snR10, U3, and U14, which cluster in the expansion segments of the 18S rRNA. Our data suggest that these snoRNAs bridge interactions between the expansion segments, thereby forming an extensive interaction network that likely promotes pre-rRNA maturation and folding in early pre-ribosomal complexes and establishes long-range rRNA interactions during ribosome synthesis.

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Author:Roman Martin, Philipp Hackert, Maike Ruprecht, Stefan Simm, Lukas Brüning, Oliver Mirus, Katherine E. Sloan, Grzegorz Kudla, Enrico SchleiffORCiDGND, Markus T. Bohnsack
URN:urn:nbn:de:hebis:30:3-344182
DOI:https://doi.org/10.1261/rna.044669.114
ISSN:1469-9001
ISSN:1355-8382
Pubmed Id:https://pubmed.ncbi.nlm.nih.gov/24947498
Parent Title (English):RNA
Publisher:Laboratory Press
Place of publication:Cold Spring Harbor, NY
Document Type:Article
Language:English
Date of Publication (online):2014/06/19
Date of first Publication:2014/06/19
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2014/07/24
Tag:RNA helicase; RNA–protein complex; ribosome biogenesis; small nucleolar RNA
Volume:20
Issue:8
Page Number:10
First Page:1173
Last Page:1182
Note:
This article, published in RNA, is available under a Creative Commons License (Attribution 4.0 International), as described at http://creativecommons.org/licenses/by/4.0/.
HeBIS-PPN:365993824
Institutes:Biowissenschaften / Biowissenschaften
Exzellenzcluster / Exzellenzcluster Makromolekulare Komplexe
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie
Sammlungen:Universitätspublikationen
Sammlung Biologie / Sondersammelgebiets-Volltexte
Licence (German):License LogoCreative Commons - Namensnennung 4.0