Differential Centrifugation to Enrich Bacterial Ribonucleoprotein Bodies (BR bodies) from Caulobacter crescentus

Bacterial RNP bodies (BR bodies) contain the mRNA decay machinery, but the collection of associated RNAs and proteins are poorly defined. Here, we present a protocol for the rapid differential centrifugation-based enrichment of BR bodies from Caulobacter crescentus cells. As native BR bodies are hig...

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Autores principales: Muthunayake, Nisansala S., Al-Husini, Nadra, Schrader, Jared M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7757671/
https://www.ncbi.nlm.nih.gov/pubmed/33377099
http://dx.doi.org/10.1016/j.xpro.2020.100205
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author Muthunayake, Nisansala S.
Al-Husini, Nadra
Schrader, Jared M.
author_facet Muthunayake, Nisansala S.
Al-Husini, Nadra
Schrader, Jared M.
author_sort Muthunayake, Nisansala S.
collection PubMed
description Bacterial RNP bodies (BR bodies) contain the mRNA decay machinery, but the collection of associated RNAs and proteins are poorly defined. Here, we present a protocol for the rapid differential centrifugation-based enrichment of BR bodies from Caulobacter crescentus cells. As native BR bodies are highly labile and dissociate by degrading internal mRNAs, an active site mutant of RNase E, which blocks dissolution of BR bodies, allows BR-body stabilization during enrichment. For complete details on the use and execution of this protocol, please refer to Al-Husini et al. (2020).
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spelling pubmed-77576712020-12-28 Differential Centrifugation to Enrich Bacterial Ribonucleoprotein Bodies (BR bodies) from Caulobacter crescentus Muthunayake, Nisansala S. Al-Husini, Nadra Schrader, Jared M. STAR Protoc Protocol Bacterial RNP bodies (BR bodies) contain the mRNA decay machinery, but the collection of associated RNAs and proteins are poorly defined. Here, we present a protocol for the rapid differential centrifugation-based enrichment of BR bodies from Caulobacter crescentus cells. As native BR bodies are highly labile and dissociate by degrading internal mRNAs, an active site mutant of RNase E, which blocks dissolution of BR bodies, allows BR-body stabilization during enrichment. For complete details on the use and execution of this protocol, please refer to Al-Husini et al. (2020). Elsevier 2020-12-09 /pmc/articles/PMC7757671/ /pubmed/33377099 http://dx.doi.org/10.1016/j.xpro.2020.100205 Text en © 2020 The Author(s) http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Muthunayake, Nisansala S.
Al-Husini, Nadra
Schrader, Jared M.
Differential Centrifugation to Enrich Bacterial Ribonucleoprotein Bodies (BR bodies) from Caulobacter crescentus
title Differential Centrifugation to Enrich Bacterial Ribonucleoprotein Bodies (BR bodies) from Caulobacter crescentus
title_full Differential Centrifugation to Enrich Bacterial Ribonucleoprotein Bodies (BR bodies) from Caulobacter crescentus
title_fullStr Differential Centrifugation to Enrich Bacterial Ribonucleoprotein Bodies (BR bodies) from Caulobacter crescentus
title_full_unstemmed Differential Centrifugation to Enrich Bacterial Ribonucleoprotein Bodies (BR bodies) from Caulobacter crescentus
title_short Differential Centrifugation to Enrich Bacterial Ribonucleoprotein Bodies (BR bodies) from Caulobacter crescentus
title_sort differential centrifugation to enrich bacterial ribonucleoprotein bodies (br bodies) from caulobacter crescentus
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7757671/
https://www.ncbi.nlm.nih.gov/pubmed/33377099
http://dx.doi.org/10.1016/j.xpro.2020.100205
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