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Single-molecule FRET unmasks structural subpopulations and crucial molecular events during FUS low-complexity domain phase separation
Biomolecular condensates formed via phase separation of proteins and nucleic acids are thought to be associated with a wide range of cellular functions and dysfunctions. We dissect critical molecular events associated with phase separation of an intrinsically disordered prion-like low-complexity dom...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10643395/ https://www.ncbi.nlm.nih.gov/pubmed/37957147 http://dx.doi.org/10.1038/s41467-023-43225-y |
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author | Joshi, Ashish Walimbe, Anuja Avni, Anamika Rai, Sandeep K. Arora, Lisha Sarkar, Snehasis Mukhopadhyay, Samrat |
author_facet | Joshi, Ashish Walimbe, Anuja Avni, Anamika Rai, Sandeep K. Arora, Lisha Sarkar, Snehasis Mukhopadhyay, Samrat |
author_sort | Joshi, Ashish |
collection | PubMed |
description | Biomolecular condensates formed via phase separation of proteins and nucleic acids are thought to be associated with a wide range of cellular functions and dysfunctions. We dissect critical molecular events associated with phase separation of an intrinsically disordered prion-like low-complexity domain of Fused in Sarcoma by performing single-molecule studies permitting us to access the wealth of molecular information that is skewed in conventional ensemble experiments. Our single-molecule FRET experiments reveal the coexistence of two conformationally distinct subpopulations in the monomeric form. Single-droplet single-molecule FRET studies coupled with fluorescence correlation spectroscopy, picosecond time-resolved fluorescence anisotropy, and vibrational Raman spectroscopy indicate that structural unwinding switches intramolecular interactions into intermolecular contacts allowing the formation of a dynamic network within condensates. A disease-related mutation introduces enhanced structural plasticity engendering greater interchain interactions that can accelerate pathological aggregation. Our findings provide key mechanistic underpinnings of sequence-encoded dynamically-controlled structural unzipping resulting in biological phase separation. |
format | Online Article Text |
id | pubmed-10643395 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-106433952023-11-13 Single-molecule FRET unmasks structural subpopulations and crucial molecular events during FUS low-complexity domain phase separation Joshi, Ashish Walimbe, Anuja Avni, Anamika Rai, Sandeep K. Arora, Lisha Sarkar, Snehasis Mukhopadhyay, Samrat Nat Commun Article Biomolecular condensates formed via phase separation of proteins and nucleic acids are thought to be associated with a wide range of cellular functions and dysfunctions. We dissect critical molecular events associated with phase separation of an intrinsically disordered prion-like low-complexity domain of Fused in Sarcoma by performing single-molecule studies permitting us to access the wealth of molecular information that is skewed in conventional ensemble experiments. Our single-molecule FRET experiments reveal the coexistence of two conformationally distinct subpopulations in the monomeric form. Single-droplet single-molecule FRET studies coupled with fluorescence correlation spectroscopy, picosecond time-resolved fluorescence anisotropy, and vibrational Raman spectroscopy indicate that structural unwinding switches intramolecular interactions into intermolecular contacts allowing the formation of a dynamic network within condensates. A disease-related mutation introduces enhanced structural plasticity engendering greater interchain interactions that can accelerate pathological aggregation. Our findings provide key mechanistic underpinnings of sequence-encoded dynamically-controlled structural unzipping resulting in biological phase separation. Nature Publishing Group UK 2023-11-13 /pmc/articles/PMC10643395/ /pubmed/37957147 http://dx.doi.org/10.1038/s41467-023-43225-y Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Joshi, Ashish Walimbe, Anuja Avni, Anamika Rai, Sandeep K. Arora, Lisha Sarkar, Snehasis Mukhopadhyay, Samrat Single-molecule FRET unmasks structural subpopulations and crucial molecular events during FUS low-complexity domain phase separation |
title | Single-molecule FRET unmasks structural subpopulations and crucial molecular events during FUS low-complexity domain phase separation |
title_full | Single-molecule FRET unmasks structural subpopulations and crucial molecular events during FUS low-complexity domain phase separation |
title_fullStr | Single-molecule FRET unmasks structural subpopulations and crucial molecular events during FUS low-complexity domain phase separation |
title_full_unstemmed | Single-molecule FRET unmasks structural subpopulations and crucial molecular events during FUS low-complexity domain phase separation |
title_short | Single-molecule FRET unmasks structural subpopulations and crucial molecular events during FUS low-complexity domain phase separation |
title_sort | single-molecule fret unmasks structural subpopulations and crucial molecular events during fus low-complexity domain phase separation |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10643395/ https://www.ncbi.nlm.nih.gov/pubmed/37957147 http://dx.doi.org/10.1038/s41467-023-43225-y |
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