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Light activation of transcription: photocaging of nucleotides for control over RNA polymerization

We describe the use of ATP caged with [7-(diethylamino)coumarin-4-yl]methyl (DEACM) for light-controlled in vitro transcription reactions. Polymerization is blocked when DEACM is bonded to the gamma phosphate group of the ATP molecule. Controlled light irradiation releases ATP and transcription is i...

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Detalles Bibliográficos
Autores principales: Pinheiro, André Vidal, Baptista, Pedro, Lima, João Carlos
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2504310/
https://www.ncbi.nlm.nih.gov/pubmed/18586819
http://dx.doi.org/10.1093/nar/gkn415
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author Pinheiro, André Vidal
Baptista, Pedro
Lima, João Carlos
author_facet Pinheiro, André Vidal
Baptista, Pedro
Lima, João Carlos
author_sort Pinheiro, André Vidal
collection PubMed
description We describe the use of ATP caged with [7-(diethylamino)coumarin-4-yl]methyl (DEACM) for light-controlled in vitro transcription reactions. Polymerization is blocked when DEACM is bonded to the gamma phosphate group of the ATP molecule. Controlled light irradiation releases ATP and transcription is initiated. In order to provide full control over the process, conditions involved in substrate release, nucleotide availability after release and the effect of the released coumarin in RNA polymerization were assessed in further detail. Together, our data provide the first direct evidence of control over enzymatic polymerization of nucleic acids through light. This approach may provide researchers with a unique tool for the study of biological processes at a molecular level.
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spelling pubmed-25043102008-08-08 Light activation of transcription: photocaging of nucleotides for control over RNA polymerization Pinheiro, André Vidal Baptista, Pedro Lima, João Carlos Nucleic Acids Res Methods Online We describe the use of ATP caged with [7-(diethylamino)coumarin-4-yl]methyl (DEACM) for light-controlled in vitro transcription reactions. Polymerization is blocked when DEACM is bonded to the gamma phosphate group of the ATP molecule. Controlled light irradiation releases ATP and transcription is initiated. In order to provide full control over the process, conditions involved in substrate release, nucleotide availability after release and the effect of the released coumarin in RNA polymerization were assessed in further detail. Together, our data provide the first direct evidence of control over enzymatic polymerization of nucleic acids through light. This approach may provide researchers with a unique tool for the study of biological processes at a molecular level. Oxford University Press 2008-08 2008-06-27 /pmc/articles/PMC2504310/ /pubmed/18586819 http://dx.doi.org/10.1093/nar/gkn415 Text en © 2008 The Author(s) http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Pinheiro, André Vidal
Baptista, Pedro
Lima, João Carlos
Light activation of transcription: photocaging of nucleotides for control over RNA polymerization
title Light activation of transcription: photocaging of nucleotides for control over RNA polymerization
title_full Light activation of transcription: photocaging of nucleotides for control over RNA polymerization
title_fullStr Light activation of transcription: photocaging of nucleotides for control over RNA polymerization
title_full_unstemmed Light activation of transcription: photocaging of nucleotides for control over RNA polymerization
title_short Light activation of transcription: photocaging of nucleotides for control over RNA polymerization
title_sort light activation of transcription: photocaging of nucleotides for control over rna polymerization
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2504310/
https://www.ncbi.nlm.nih.gov/pubmed/18586819
http://dx.doi.org/10.1093/nar/gkn415
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