Switching transcription with bacterial RNA polymerase through photocaging, photorelease and phosphorylation reactions in the major groove of DNA

We report proof of principle biomimetic switching of transcription in vitro through non-natural chemical reactions in the major groove of DNA templates. Photocaged DNA templates containing nitrobenzyl-protected 5-hydroxymethyluracil or – cytosine permitted no transcription with E. coli RNA polymeras...

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Autores principales: Vaníková, Zuzana, Janoušková, Martina, Kambová, Milada, Krásný, Libor, Hocek, Michal
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Royal Society of Chemistry 2019
Materias:
Chemistry
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6457204/
https://www.ncbi.nlm.nih.gov/pubmed/31015933
http://dx.doi.org/10.1039/c9sc00205g
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author Vaníková, Zuzana
Janoušková, Martina
Kambová, Milada
Krásný, Libor
Hocek, Michal
author_facet Vaníková, Zuzana
Janoušková, Martina
Kambová, Milada
Krásný, Libor
Hocek, Michal
author_sort Vaníková, Zuzana
collection PubMed
description We report proof of principle biomimetic switching of transcription in vitro through non-natural chemical reactions in the major groove of DNA templates. Photocaged DNA templates containing nitrobenzyl-protected 5-hydroxymethyluracil or – cytosine permitted no transcription with E. coli RNA polymerase (OFF state). Their irradiation with 400 nm light resulted in DNA templates containing hydroxymethylpyrimidines, which switched transcription ON with a higher yield (250–350%) compared to non-modified DNA. Phosphorylation of templates containing 5-hydroxymethyluracil (but not 5-hydroxymethylcytosine) then turned transcription OFF again. It is the first step towards artificial bioorthogonal chemical epigenetics.
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spelling pubmed-64572042019-04-23 Switching transcription with bacterial RNA polymerase through photocaging, photorelease and phosphorylation reactions in the major groove of DNA Vaníková, Zuzana Janoušková, Martina Kambová, Milada Krásný, Libor Hocek, Michal Chem Sci Chemistry We report proof of principle biomimetic switching of transcription in vitro through non-natural chemical reactions in the major groove of DNA templates. Photocaged DNA templates containing nitrobenzyl-protected 5-hydroxymethyluracil or – cytosine permitted no transcription with E. coli RNA polymerase (OFF state). Their irradiation with 400 nm light resulted in DNA templates containing hydroxymethylpyrimidines, which switched transcription ON with a higher yield (250–350%) compared to non-modified DNA. Phosphorylation of templates containing 5-hydroxymethyluracil (but not 5-hydroxymethylcytosine) then turned transcription OFF again. It is the first step towards artificial bioorthogonal chemical epigenetics. Royal Society of Chemistry 2019-03-04 /pmc/articles/PMC6457204/ /pubmed/31015933 http://dx.doi.org/10.1039/c9sc00205g Text en This journal is © The Royal Society of Chemistry 2019 http://creativecommons.org/licenses/by/3.0/ This article is freely available. This article is licensed under a Creative Commons Attribution 3.0 Unported Licence (CC BY 3.0)
spellingShingle Chemistry
Vaníková, Zuzana
Janoušková, Martina
Kambová, Milada
Krásný, Libor
Hocek, Michal
Switching transcription with bacterial RNA polymerase through photocaging, photorelease and phosphorylation reactions in the major groove of DNA
title Switching transcription with bacterial RNA polymerase through photocaging, photorelease and phosphorylation reactions in the major groove of DNA
title_full Switching transcription with bacterial RNA polymerase through photocaging, photorelease and phosphorylation reactions in the major groove of DNA
title_fullStr Switching transcription with bacterial RNA polymerase through photocaging, photorelease and phosphorylation reactions in the major groove of DNA
title_full_unstemmed Switching transcription with bacterial RNA polymerase through photocaging, photorelease and phosphorylation reactions in the major groove of DNA
title_short Switching transcription with bacterial RNA polymerase through photocaging, photorelease and phosphorylation reactions in the major groove of DNA
title_sort switching transcription with bacterial rna polymerase through photocaging, photorelease and phosphorylation reactions in the major groove of dna
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6457204/
https://www.ncbi.nlm.nih.gov/pubmed/31015933
http://dx.doi.org/10.1039/c9sc00205g
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