RNA-seq analyses reveal the relevance of RNAs involved in ribosomal complex to induce mammalian prion protein aggregation and phase separation in vitro

Conformational conversion of cellular prion protein (PrPC) into infectious PrP (PrPSc) is one of the most intriguing processes in modern Biology. It is well accepted that this transition is catalysed by one or more cofactors that lower the energy barrier between the different PrP forms. Among potent...

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Main Authors: Ana C. Tahira, Mariana P. B. Gomes, Maria Heloisa Freire, Marcelly Muxfeldt, Francisco Prosdocimi, Yulli M. Passos, Murilo Sena Amaral, Leticia P. Felix Valadão, Luciana P. Rangel, Jerson L. Silva, Sergio Verjovski-Almeida, Yraima Cordeiro
Format: Article
Language:English
Published: Taylor & Francis Group 2025-12-01
Series:RNA Biology
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Online Access:https://www.tandfonline.com/doi/10.1080/15476286.2025.2508107
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author Ana C. Tahira
Mariana P. B. Gomes
Maria Heloisa Freire
Marcelly Muxfeldt
Francisco Prosdocimi
Yulli M. Passos
Murilo Sena Amaral
Leticia P. Felix Valadão
Luciana P. Rangel
Jerson L. Silva
Sergio Verjovski-Almeida
Yraima Cordeiro
author_facet Ana C. Tahira
Mariana P. B. Gomes
Maria Heloisa Freire
Marcelly Muxfeldt
Francisco Prosdocimi
Yulli M. Passos
Murilo Sena Amaral
Leticia P. Felix Valadão
Luciana P. Rangel
Jerson L. Silva
Sergio Verjovski-Almeida
Yraima Cordeiro
author_sort Ana C. Tahira
collection DOAJ
description Conformational conversion of cellular prion protein (PrPC) into infectious PrP (PrPSc) is one of the most intriguing processes in modern Biology. It is well accepted that this transition is catalysed by one or more cofactors that lower the energy barrier between the different PrP forms. Among potential candidates, RNA molecules are strong contenders. Our group has pursued nucleic acids, both DNA and RNA, capable of inducing PrP misfolding, aggregation, and, more recently, phase separation, a process proposed to precede aggregation in degenerative disorders. We found that the interaction between recombinant PrP (rPrP) and total RNA extracted from neuroblastoma cells (N2aRNA) results in significant structural alterations. Here, we use rPrP:N2aRNA as a model to search for RNAs capable of inducing full-length murine rPrP phase separation and/or aggregation. N2aRNA was incubated with rPrP and after that, RNA-seq analysis was conducted with RNAs isolated from the insoluble material using two different protocols. We analysed thousands of RNA-seq reads, most of which represented ribosomal RNA molecules. The set of recovered molecules is heterogeneous; nevertheless, three low-complexity consensus motifs within the sequences of RNAs involved in ribosomal complex were identified as significantly enriched in the RNAs bound to rPrP, suggesting that a population of RNAs is responsible for inducing PrP phase transitions. We hypothesize that RNA transcripts enriched in a set of low complexity motif sequences with predicted structural similarities can be involved in PrPC binding. This interaction would lead to phase separation and, ultimately, result in aggregation into scrapie-like species, in a stoichiometry-dependent manner.
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spelling doaj-art-bf3b8cd2a16b4adfad3b5ddd383c80452025-08-20T02:17:01ZengTaylor & Francis GroupRNA Biology1547-62861555-85842025-12-0122111610.1080/15476286.2025.2508107RNA-seq analyses reveal the relevance of RNAs involved in ribosomal complex to induce mammalian prion protein aggregation and phase separation in vitroAna C. Tahira0Mariana P. B. Gomes1Maria Heloisa Freire2Marcelly Muxfeldt3Francisco Prosdocimi4Yulli M. Passos5Murilo Sena Amaral6Leticia P. Felix Valadão7Luciana P. Rangel8Jerson L. Silva9Sergio Verjovski-Almeida10Yraima Cordeiro11Laboratório de Ciclo Celular, Instituto Butantan, São Paulo, BrasilFaculdade de Farmácia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, BrasilFaculdade de Farmácia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, BrasilFaculdade de Farmácia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, BrasilInstituto de Bioquímica Médica Leopoldo de Meis, Universidade Federal do Rio de Janeiro, Rio de Janeiro, BrasilFaculdade de Farmácia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, BrasilLaboratório de Ciclo Celular, Instituto Butantan, São Paulo, BrasilFaculdade de Farmácia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, BrasilFaculdade de Farmácia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, BrasilInstituto de Bioquímica Médica Leopoldo de Meis, Universidade Federal do Rio de Janeiro, Rio de Janeiro, BrasilLaboratório de Ciclo Celular, Instituto Butantan, São Paulo, BrasilFaculdade de Farmácia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, BrasilConformational conversion of cellular prion protein (PrPC) into infectious PrP (PrPSc) is one of the most intriguing processes in modern Biology. It is well accepted that this transition is catalysed by one or more cofactors that lower the energy barrier between the different PrP forms. Among potential candidates, RNA molecules are strong contenders. Our group has pursued nucleic acids, both DNA and RNA, capable of inducing PrP misfolding, aggregation, and, more recently, phase separation, a process proposed to precede aggregation in degenerative disorders. We found that the interaction between recombinant PrP (rPrP) and total RNA extracted from neuroblastoma cells (N2aRNA) results in significant structural alterations. Here, we use rPrP:N2aRNA as a model to search for RNAs capable of inducing full-length murine rPrP phase separation and/or aggregation. N2aRNA was incubated with rPrP and after that, RNA-seq analysis was conducted with RNAs isolated from the insoluble material using two different protocols. We analysed thousands of RNA-seq reads, most of which represented ribosomal RNA molecules. The set of recovered molecules is heterogeneous; nevertheless, three low-complexity consensus motifs within the sequences of RNAs involved in ribosomal complex were identified as significantly enriched in the RNAs bound to rPrP, suggesting that a population of RNAs is responsible for inducing PrP phase transitions. We hypothesize that RNA transcripts enriched in a set of low complexity motif sequences with predicted structural similarities can be involved in PrPC binding. This interaction would lead to phase separation and, ultimately, result in aggregation into scrapie-like species, in a stoichiometry-dependent manner.https://www.tandfonline.com/doi/10.1080/15476286.2025.2508107Prion proteinRNA-seqChIP-seqprion diseasesRNAnucleic acid
spellingShingle Ana C. Tahira
Mariana P. B. Gomes
Maria Heloisa Freire
Marcelly Muxfeldt
Francisco Prosdocimi
Yulli M. Passos
Murilo Sena Amaral
Leticia P. Felix Valadão
Luciana P. Rangel
Jerson L. Silva
Sergio Verjovski-Almeida
Yraima Cordeiro
RNA-seq analyses reveal the relevance of RNAs involved in ribosomal complex to induce mammalian prion protein aggregation and phase separation in vitro
RNA Biology
Prion protein
RNA-seq
ChIP-seq
prion diseases
RNA
nucleic acid
title RNA-seq analyses reveal the relevance of RNAs involved in ribosomal complex to induce mammalian prion protein aggregation and phase separation in vitro
title_full RNA-seq analyses reveal the relevance of RNAs involved in ribosomal complex to induce mammalian prion protein aggregation and phase separation in vitro
title_fullStr RNA-seq analyses reveal the relevance of RNAs involved in ribosomal complex to induce mammalian prion protein aggregation and phase separation in vitro
title_full_unstemmed RNA-seq analyses reveal the relevance of RNAs involved in ribosomal complex to induce mammalian prion protein aggregation and phase separation in vitro
title_short RNA-seq analyses reveal the relevance of RNAs involved in ribosomal complex to induce mammalian prion protein aggregation and phase separation in vitro
title_sort rna seq analyses reveal the relevance of rnas involved in ribosomal complex to induce mammalian prion protein aggregation and phase separation in vitro
topic Prion protein
RNA-seq
ChIP-seq
prion diseases
RNA
nucleic acid
url https://www.tandfonline.com/doi/10.1080/15476286.2025.2508107
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