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Synergistic interaction network between the snR30 RNP, Utp23, and ribosomal RNA during ribosome synthesis

snR30/U17 is a highly conserved H/ACA RNA that is required for maturation of the small ribosomal subunit in eukaryotes. By base-pairing to the expansion segment 6 (ES6) of 18S ribosomal RNA (rRNA), the snR30 H/ACA Ribonucleoprotein (RNP) indirectly facilitates processing of the precursor rRNA (pre-r...

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Autores principales: Vos, Timothy J., Kothe, Ute
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9176245/
https://www.ncbi.nlm.nih.gov/pubmed/35648701
http://dx.doi.org/10.1080/15476286.2022.2078092
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author Vos, Timothy J.
Kothe, Ute
author_facet Vos, Timothy J.
Kothe, Ute
author_sort Vos, Timothy J.
collection PubMed
description snR30/U17 is a highly conserved H/ACA RNA that is required for maturation of the small ribosomal subunit in eukaryotes. By base-pairing to the expansion segment 6 (ES6) of 18S ribosomal RNA (rRNA), the snR30 H/ACA Ribonucleoprotein (RNP) indirectly facilitates processing of the precursor rRNA (pre-rRNA) together with other proteins such as Utp23 and other RNAs acting as ribosome assembly factors. However, the details of the molecular interaction network of snR30 and its binding partners and how these interactions contribute to pre-rRNA processing remains unknown. Here, we report the in vitro reconstitution of a Saccharomyces cerevisiae snR30 RNP and quantitative characterization of the interactions of snR30, H/ACA proteins, the Utp23 protein and ES6 of the 18S rRNA. The snR30 RNA is bound tightly by both H/ACA proteins and Utp23. We dissected the importance of different 18S rRNA regions for snR30 RNP binding and demonstrated that the snR30 complex is tightly anchored on the pre-rRNA through base-pairing to ES6 whereas other reported rRNA binding sites do not contribute to the affinity of the snR30 RNP. On its own, the ribosome assembly factor Utp23 binds in a tight, but unspecific manner to RNA. However, in complex with the snR30 RNP, Utp23 increases the affinity of the RNP for rRNA revealing synergies between snR30 RNP and Utp23 which are enhancing specificity and affinity for rRNA, respectively. Together, these findings provide mechanistic insights how the snR30 RNP and Utp23 cooperate to interact tightly and specifically with rRNA during the early stages of ribosome biogenesis.
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spelling pubmed-91762452022-06-09 Synergistic interaction network between the snR30 RNP, Utp23, and ribosomal RNA during ribosome synthesis Vos, Timothy J. Kothe, Ute RNA Biol Research Paper snR30/U17 is a highly conserved H/ACA RNA that is required for maturation of the small ribosomal subunit in eukaryotes. By base-pairing to the expansion segment 6 (ES6) of 18S ribosomal RNA (rRNA), the snR30 H/ACA Ribonucleoprotein (RNP) indirectly facilitates processing of the precursor rRNA (pre-rRNA) together with other proteins such as Utp23 and other RNAs acting as ribosome assembly factors. However, the details of the molecular interaction network of snR30 and its binding partners and how these interactions contribute to pre-rRNA processing remains unknown. Here, we report the in vitro reconstitution of a Saccharomyces cerevisiae snR30 RNP and quantitative characterization of the interactions of snR30, H/ACA proteins, the Utp23 protein and ES6 of the 18S rRNA. The snR30 RNA is bound tightly by both H/ACA proteins and Utp23. We dissected the importance of different 18S rRNA regions for snR30 RNP binding and demonstrated that the snR30 complex is tightly anchored on the pre-rRNA through base-pairing to ES6 whereas other reported rRNA binding sites do not contribute to the affinity of the snR30 RNP. On its own, the ribosome assembly factor Utp23 binds in a tight, but unspecific manner to RNA. However, in complex with the snR30 RNP, Utp23 increases the affinity of the RNP for rRNA revealing synergies between snR30 RNP and Utp23 which are enhancing specificity and affinity for rRNA, respectively. Together, these findings provide mechanistic insights how the snR30 RNP and Utp23 cooperate to interact tightly and specifically with rRNA during the early stages of ribosome biogenesis. Taylor & Francis 2022-06-01 /pmc/articles/PMC9176245/ /pubmed/35648701 http://dx.doi.org/10.1080/15476286.2022.2078092 Text en © 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Paper
Vos, Timothy J.
Kothe, Ute
Synergistic interaction network between the snR30 RNP, Utp23, and ribosomal RNA during ribosome synthesis
title Synergistic interaction network between the snR30 RNP, Utp23, and ribosomal RNA during ribosome synthesis
title_full Synergistic interaction network between the snR30 RNP, Utp23, and ribosomal RNA during ribosome synthesis
title_fullStr Synergistic interaction network between the snR30 RNP, Utp23, and ribosomal RNA during ribosome synthesis
title_full_unstemmed Synergistic interaction network between the snR30 RNP, Utp23, and ribosomal RNA during ribosome synthesis
title_short Synergistic interaction network between the snR30 RNP, Utp23, and ribosomal RNA during ribosome synthesis
title_sort synergistic interaction network between the snr30 rnp, utp23, and ribosomal rna during ribosome synthesis
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9176245/
https://www.ncbi.nlm.nih.gov/pubmed/35648701
http://dx.doi.org/10.1080/15476286.2022.2078092
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