Protocol for an in vitro assay to study HIV-1 Tat methylation

A critical virus-encoded regulator of HIV-1 transcription is the Tat protein, which is required to potently activate transcription. Tat is regulated by a wide variety of post-translational modifications. This protocol describes an in vitro assay to study Tat methylation. We describe steps for incorp...

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Autores principales: Boehm, Daniela, Kaehlcke, Katrin, Schnoelzer, Martina, Ott, Melanie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10694577/
https://www.ncbi.nlm.nih.gov/pubmed/37979180
http://dx.doi.org/10.1016/j.xpro.2023.102687
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author Boehm, Daniela
Kaehlcke, Katrin
Schnoelzer, Martina
Ott, Melanie
author_facet Boehm, Daniela
Kaehlcke, Katrin
Schnoelzer, Martina
Ott, Melanie
author_sort Boehm, Daniela
collection PubMed
description A critical virus-encoded regulator of HIV-1 transcription is the Tat protein, which is required to potently activate transcription. Tat is regulated by a wide variety of post-translational modifications. This protocol describes an in vitro assay to study Tat methylation. We describe steps for incorporation of radioactive methyl groups into Tat protein, visualization by gel analysis, Coomassie blue stain, gel drying, and detection by autoradiography. This protocol can also be used to assess methylation in other proteins such as histones. For complete details on the use and execution of this protocol, please refer to Boehm et al. (2023).(1)
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spelling pubmed-106945772023-12-05 Protocol for an in vitro assay to study HIV-1 Tat methylation Boehm, Daniela Kaehlcke, Katrin Schnoelzer, Martina Ott, Melanie STAR Protoc Protocol A critical virus-encoded regulator of HIV-1 transcription is the Tat protein, which is required to potently activate transcription. Tat is regulated by a wide variety of post-translational modifications. This protocol describes an in vitro assay to study Tat methylation. We describe steps for incorporation of radioactive methyl groups into Tat protein, visualization by gel analysis, Coomassie blue stain, gel drying, and detection by autoradiography. This protocol can also be used to assess methylation in other proteins such as histones. For complete details on the use and execution of this protocol, please refer to Boehm et al. (2023).(1) Elsevier 2023-11-18 /pmc/articles/PMC10694577/ /pubmed/37979180 http://dx.doi.org/10.1016/j.xpro.2023.102687 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Boehm, Daniela
Kaehlcke, Katrin
Schnoelzer, Martina
Ott, Melanie
Protocol for an in vitro assay to study HIV-1 Tat methylation
title Protocol for an in vitro assay to study HIV-1 Tat methylation
title_full Protocol for an in vitro assay to study HIV-1 Tat methylation
title_fullStr Protocol for an in vitro assay to study HIV-1 Tat methylation
title_full_unstemmed Protocol for an in vitro assay to study HIV-1 Tat methylation
title_short Protocol for an in vitro assay to study HIV-1 Tat methylation
title_sort protocol for an in vitro assay to study hiv-1 tat methylation
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10694577/
https://www.ncbi.nlm.nih.gov/pubmed/37979180
http://dx.doi.org/10.1016/j.xpro.2023.102687
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