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Mesoscale structure–function relationships in mitochondrial transcriptional condensates

In live cells, phase separation is thought to organize macromolecules into membraneless structures known as biomolecular condensates. Here, we reconstituted transcription in condensates from purified mitochondrial components using optimized in vitro reaction conditions to probe the structure–functio...

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Autores principales: Feric, Marina, Sarfallah, Azadeh, Dar, Furqan, Temiakov, Dmitry, Pappu, Rohit V., Misteli, Tom
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9565167/
https://www.ncbi.nlm.nih.gov/pubmed/36191226
http://dx.doi.org/10.1073/pnas.2207303119
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author Feric, Marina
Sarfallah, Azadeh
Dar, Furqan
Temiakov, Dmitry
Pappu, Rohit V.
Misteli, Tom
author_facet Feric, Marina
Sarfallah, Azadeh
Dar, Furqan
Temiakov, Dmitry
Pappu, Rohit V.
Misteli, Tom
author_sort Feric, Marina
collection PubMed
description In live cells, phase separation is thought to organize macromolecules into membraneless structures known as biomolecular condensates. Here, we reconstituted transcription in condensates from purified mitochondrial components using optimized in vitro reaction conditions to probe the structure–function relationships of biomolecular condensates. We find that the core components of the mt-transcription machinery form multiphasic, viscoelastic condensates in vitro. Strikingly, the rates of condensate-mediated transcription are substantially lower than in solution. The condensate-mediated decrease in transcriptional rates is associated with the formation of vesicle-like structures that are driven by the production and accumulation of RNA during transcription. The generation of RNA alters the global phase behavior and organization of transcription components within condensates. Coarse-grained simulations of mesoscale structures at equilibrium show that the components stably assemble into multiphasic condensates and that the vesicles formed in vitro are the result of dynamical arrest. Overall, our findings illustrate the complex phase behavior of transcribing, multicomponent condensates, and they highlight the intimate, bidirectional interplay of structure and function in transcriptional condensates.
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spelling pubmed-95651672023-04-03 Mesoscale structure–function relationships in mitochondrial transcriptional condensates Feric, Marina Sarfallah, Azadeh Dar, Furqan Temiakov, Dmitry Pappu, Rohit V. Misteli, Tom Proc Natl Acad Sci U S A Physical Sciences In live cells, phase separation is thought to organize macromolecules into membraneless structures known as biomolecular condensates. Here, we reconstituted transcription in condensates from purified mitochondrial components using optimized in vitro reaction conditions to probe the structure–function relationships of biomolecular condensates. We find that the core components of the mt-transcription machinery form multiphasic, viscoelastic condensates in vitro. Strikingly, the rates of condensate-mediated transcription are substantially lower than in solution. The condensate-mediated decrease in transcriptional rates is associated with the formation of vesicle-like structures that are driven by the production and accumulation of RNA during transcription. The generation of RNA alters the global phase behavior and organization of transcription components within condensates. Coarse-grained simulations of mesoscale structures at equilibrium show that the components stably assemble into multiphasic condensates and that the vesicles formed in vitro are the result of dynamical arrest. Overall, our findings illustrate the complex phase behavior of transcribing, multicomponent condensates, and they highlight the intimate, bidirectional interplay of structure and function in transcriptional condensates. National Academy of Sciences 2022-10-03 2022-10-11 /pmc/articles/PMC9565167/ /pubmed/36191226 http://dx.doi.org/10.1073/pnas.2207303119 Text en Copyright © 2022 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/This article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) .
spellingShingle Physical Sciences
Feric, Marina
Sarfallah, Azadeh
Dar, Furqan
Temiakov, Dmitry
Pappu, Rohit V.
Misteli, Tom
Mesoscale structure–function relationships in mitochondrial transcriptional condensates
title Mesoscale structure–function relationships in mitochondrial transcriptional condensates
title_full Mesoscale structure–function relationships in mitochondrial transcriptional condensates
title_fullStr Mesoscale structure–function relationships in mitochondrial transcriptional condensates
title_full_unstemmed Mesoscale structure–function relationships in mitochondrial transcriptional condensates
title_short Mesoscale structure–function relationships in mitochondrial transcriptional condensates
title_sort mesoscale structure–function relationships in mitochondrial transcriptional condensates
topic Physical Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9565167/
https://www.ncbi.nlm.nih.gov/pubmed/36191226
http://dx.doi.org/10.1073/pnas.2207303119
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