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High-fidelity reconstitution of stress granules and nucleoli in mammalian cellular lysate
Liquid–liquid phase separation (LLPS) is a mechanism of intracellular organization that underlies the assembly of a variety of RNP granules. Fundamental biophysical principles governing LLPS during granule assembly have been revealed by simple in vitro systems, but these systems have limitations whe...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Rockefeller University Press
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7845923/ https://www.ncbi.nlm.nih.gov/pubmed/33502444 http://dx.doi.org/10.1083/jcb.202009079 |
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author | Freibaum, Brian D. Messing, James Yang, Peiguo Kim, Hong Joo Taylor, J. Paul |
author_facet | Freibaum, Brian D. Messing, James Yang, Peiguo Kim, Hong Joo Taylor, J. Paul |
author_sort | Freibaum, Brian D. |
collection | PubMed |
description | Liquid–liquid phase separation (LLPS) is a mechanism of intracellular organization that underlies the assembly of a variety of RNP granules. Fundamental biophysical principles governing LLPS during granule assembly have been revealed by simple in vitro systems, but these systems have limitations when studying the biology of complex, multicomponent RNP granules. Visualization of RNP granules in cells has validated key principles revealed by simple in vitro systems, but this approach presents difficulties for interrogating biophysical features of RNP granules and provides limited ability to manipulate protein, nucleic acid, or small molecule concentrations. Here, we introduce a system that builds upon recent insights into the mechanisms underlying RNP granule assembly and permits high-fidelity reconstitution of stress granules and the granular component of nucleoli in mammalian cellular lysate. This system fills the gap between simple in vitro systems and live cells and allows for a variety of studies of membraneless organelles, including the development of therapeutics that modify properties of specific condensates. |
format | Online Article Text |
id | pubmed-7845923 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-78459232021-09-01 High-fidelity reconstitution of stress granules and nucleoli in mammalian cellular lysate Freibaum, Brian D. Messing, James Yang, Peiguo Kim, Hong Joo Taylor, J. Paul J Cell Biol Tools Liquid–liquid phase separation (LLPS) is a mechanism of intracellular organization that underlies the assembly of a variety of RNP granules. Fundamental biophysical principles governing LLPS during granule assembly have been revealed by simple in vitro systems, but these systems have limitations when studying the biology of complex, multicomponent RNP granules. Visualization of RNP granules in cells has validated key principles revealed by simple in vitro systems, but this approach presents difficulties for interrogating biophysical features of RNP granules and provides limited ability to manipulate protein, nucleic acid, or small molecule concentrations. Here, we introduce a system that builds upon recent insights into the mechanisms underlying RNP granule assembly and permits high-fidelity reconstitution of stress granules and the granular component of nucleoli in mammalian cellular lysate. This system fills the gap between simple in vitro systems and live cells and allows for a variety of studies of membraneless organelles, including the development of therapeutics that modify properties of specific condensates. Rockefeller University Press 2021-01-27 /pmc/articles/PMC7845923/ /pubmed/33502444 http://dx.doi.org/10.1083/jcb.202009079 Text en © 2021 Freibaum et al. http://www.rupress.org/terms/https://creativecommons.org/licenses/by-nc-sa/4.0/This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/). |
spellingShingle | Tools Freibaum, Brian D. Messing, James Yang, Peiguo Kim, Hong Joo Taylor, J. Paul High-fidelity reconstitution of stress granules and nucleoli in mammalian cellular lysate |
title | High-fidelity reconstitution of stress granules and nucleoli in mammalian cellular lysate |
title_full | High-fidelity reconstitution of stress granules and nucleoli in mammalian cellular lysate |
title_fullStr | High-fidelity reconstitution of stress granules and nucleoli in mammalian cellular lysate |
title_full_unstemmed | High-fidelity reconstitution of stress granules and nucleoli in mammalian cellular lysate |
title_short | High-fidelity reconstitution of stress granules and nucleoli in mammalian cellular lysate |
title_sort | high-fidelity reconstitution of stress granules and nucleoli in mammalian cellular lysate |
topic | Tools |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7845923/ https://www.ncbi.nlm.nih.gov/pubmed/33502444 http://dx.doi.org/10.1083/jcb.202009079 |
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