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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...
Autores principales: | , , |
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Formato: | Texto |
Lenguaje: | English |
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Oxford University Press
2008
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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. |
format | Text |
id | pubmed-2504310 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
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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