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Establishment of 5′–3′ interactions in mRNA independent of a continuous ribose-phosphate backbone

Functions of eukaryotic mRNAs are characterized by intramolecular interactions between their ends. We have addressed the question whether 5′ and 3′ ends meet by diffusion-controlled encounter “through solution” or by a mechanism involving the RNA backbone. For this purpose, we used a translation sys...

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Detalles Bibliográficos
Autores principales: Kluge, Florian, Götze, Michael, Wahle, Elmar
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7161349/
https://www.ncbi.nlm.nih.gov/pubmed/32111664
http://dx.doi.org/10.1261/rna.073759.119
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author Kluge, Florian
Götze, Michael
Wahle, Elmar
author_facet Kluge, Florian
Götze, Michael
Wahle, Elmar
author_sort Kluge, Florian
collection PubMed
description Functions of eukaryotic mRNAs are characterized by intramolecular interactions between their ends. We have addressed the question whether 5′ and 3′ ends meet by diffusion-controlled encounter “through solution” or by a mechanism involving the RNA backbone. For this purpose, we used a translation system derived from Drosophila embryos that displays two types of 5′–3′ interactions: Cap-dependent translation initiation is stimulated by the poly(A) tail and inhibited by Smaug recognition elements (SREs) in the 3′ UTR. Chimeric RNAs were made consisting of one RNA molecule carrying a luciferase coding sequence and a second molecule containing SREs and a poly(A) tail; the two were connected via a protein linker. The poly(A) tail stimulated translation of such chimeras even when disruption of the RNA backbone was combined with an inversion of the 5′–3′ polarity between the open reading frame and poly(A) segment. Stimulation by the poly(A) tail also decreased with increasing RNA length. Both observations suggest that contacts between the poly(A) tail and the 5′ end are established through solution, independently of the RNA backbone. In the same chimeric constructs, SRE-dependent inhibition of translation was also insensitive to disruption of the RNA backbone. Thus, tracking of the backbone is not involved in the repression of cap-dependent initiation. However, SRE-dependent repression was insensitive to mRNA length, suggesting that the contact between the SREs in the 3′ UTR and the 5′ end of the RNA might be established in a manner that differs from the contact between the poly(A) tail and the cap.
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spelling pubmed-71613492021-05-01 Establishment of 5′–3′ interactions in mRNA independent of a continuous ribose-phosphate backbone Kluge, Florian Götze, Michael Wahle, Elmar RNA Article Functions of eukaryotic mRNAs are characterized by intramolecular interactions between their ends. We have addressed the question whether 5′ and 3′ ends meet by diffusion-controlled encounter “through solution” or by a mechanism involving the RNA backbone. For this purpose, we used a translation system derived from Drosophila embryos that displays two types of 5′–3′ interactions: Cap-dependent translation initiation is stimulated by the poly(A) tail and inhibited by Smaug recognition elements (SREs) in the 3′ UTR. Chimeric RNAs were made consisting of one RNA molecule carrying a luciferase coding sequence and a second molecule containing SREs and a poly(A) tail; the two were connected via a protein linker. The poly(A) tail stimulated translation of such chimeras even when disruption of the RNA backbone was combined with an inversion of the 5′–3′ polarity between the open reading frame and poly(A) segment. Stimulation by the poly(A) tail also decreased with increasing RNA length. Both observations suggest that contacts between the poly(A) tail and the 5′ end are established through solution, independently of the RNA backbone. In the same chimeric constructs, SRE-dependent inhibition of translation was also insensitive to disruption of the RNA backbone. Thus, tracking of the backbone is not involved in the repression of cap-dependent initiation. However, SRE-dependent repression was insensitive to mRNA length, suggesting that the contact between the SREs in the 3′ UTR and the 5′ end of the RNA might be established in a manner that differs from the contact between the poly(A) tail and the cap. Cold Spring Harbor Laboratory Press 2020-05 /pmc/articles/PMC7161349/ /pubmed/32111664 http://dx.doi.org/10.1261/rna.073759.119 Text en © 2020 Kluge et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 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 Article
Kluge, Florian
Götze, Michael
Wahle, Elmar
Establishment of 5′–3′ interactions in mRNA independent of a continuous ribose-phosphate backbone
title Establishment of 5′–3′ interactions in mRNA independent of a continuous ribose-phosphate backbone
title_full Establishment of 5′–3′ interactions in mRNA independent of a continuous ribose-phosphate backbone
title_fullStr Establishment of 5′–3′ interactions in mRNA independent of a continuous ribose-phosphate backbone
title_full_unstemmed Establishment of 5′–3′ interactions in mRNA independent of a continuous ribose-phosphate backbone
title_short Establishment of 5′–3′ interactions in mRNA independent of a continuous ribose-phosphate backbone
title_sort establishment of 5′–3′ interactions in mrna independent of a continuous ribose-phosphate backbone
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7161349/
https://www.ncbi.nlm.nih.gov/pubmed/32111664
http://dx.doi.org/10.1261/rna.073759.119
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