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Methods to assess helicase and translocation activities of human nuclear RNA exosome and RNA adaptor complexes

The nuclear RNA exosome collaborates with the MTR4 helicase and RNA adaptor complexes to process, surveil, and degrade RNA. Here we outline methods to characterize RNA translocation and strand displacement by exosome-associated helicases and adaptor complexes using fluorescence-based strand displace...

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Autores principales: Puno, M. Rhyan, Lima, Christopher D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9382703/
https://www.ncbi.nlm.nih.gov/pubmed/35965016
http://dx.doi.org/10.1016/bs.mie.2022.03.060
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author Puno, M. Rhyan
Lima, Christopher D.
author_facet Puno, M. Rhyan
Lima, Christopher D.
author_sort Puno, M. Rhyan
collection PubMed
description The nuclear RNA exosome collaborates with the MTR4 helicase and RNA adaptor complexes to process, surveil, and degrade RNA. Here we outline methods to characterize RNA translocation and strand displacement by exosome-associated helicases and adaptor complexes using fluorescence-based strand displacement assays. The design and preparation of substrates suitable for analysis of helicase and decay activities of reconstituted MTR4–exosome complexes are described. To aid structural and biophysical studies, we present strategies for engineering substrates that can stall helicases during translocation, providing a means to capture snapshots of interactions and molecular steps involved in substrate translocation and delivery to the exosome.
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spelling pubmed-93827032022-08-17 Methods to assess helicase and translocation activities of human nuclear RNA exosome and RNA adaptor complexes Puno, M. Rhyan Lima, Christopher D. Methods Enzymol Article The nuclear RNA exosome collaborates with the MTR4 helicase and RNA adaptor complexes to process, surveil, and degrade RNA. Here we outline methods to characterize RNA translocation and strand displacement by exosome-associated helicases and adaptor complexes using fluorescence-based strand displacement assays. The design and preparation of substrates suitable for analysis of helicase and decay activities of reconstituted MTR4–exosome complexes are described. To aid structural and biophysical studies, we present strategies for engineering substrates that can stall helicases during translocation, providing a means to capture snapshots of interactions and molecular steps involved in substrate translocation and delivery to the exosome. 2022 2022-04-27 /pmc/articles/PMC9382703/ /pubmed/35965016 http://dx.doi.org/10.1016/bs.mie.2022.03.060 Text en https://creativecommons.org/licenses/by/4.0/This work is licensed under a Creative Commons Attribution 4.0 International License, which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format, so long as attribution is given to the creator. The license allows for commercial use.
spellingShingle Article
Puno, M. Rhyan
Lima, Christopher D.
Methods to assess helicase and translocation activities of human nuclear RNA exosome and RNA adaptor complexes
title Methods to assess helicase and translocation activities of human nuclear RNA exosome and RNA adaptor complexes
title_full Methods to assess helicase and translocation activities of human nuclear RNA exosome and RNA adaptor complexes
title_fullStr Methods to assess helicase and translocation activities of human nuclear RNA exosome and RNA adaptor complexes
title_full_unstemmed Methods to assess helicase and translocation activities of human nuclear RNA exosome and RNA adaptor complexes
title_short Methods to assess helicase and translocation activities of human nuclear RNA exosome and RNA adaptor complexes
title_sort methods to assess helicase and translocation activities of human nuclear rna exosome and rna adaptor complexes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9382703/
https://www.ncbi.nlm.nih.gov/pubmed/35965016
http://dx.doi.org/10.1016/bs.mie.2022.03.060
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