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Oxazinomycin arrests RNA polymerase at the polythymidine sequences
Oxazinomycin is a C-nucleoside antibiotic that is produced by Streptomyces hygroscopicus and closely resembles uridine. Here, we show that the oxazinomycin triphosphate is a good substrate for bacterial and eukaryotic RNA polymerases (RNAPs) and that a single incorporated oxazinomycin is rapidly ext...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6821320/ https://www.ncbi.nlm.nih.gov/pubmed/31495891 http://dx.doi.org/10.1093/nar/gkz782 |
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author | Prajapati, Ranjit K Rosenqvist, Petja Palmu, Kaisa Mäkinen, Janne J Malinen, Anssi M Virta, Pasi Metsä-Ketelä, Mikko Belogurov, Georgiy A |
author_facet | Prajapati, Ranjit K Rosenqvist, Petja Palmu, Kaisa Mäkinen, Janne J Malinen, Anssi M Virta, Pasi Metsä-Ketelä, Mikko Belogurov, Georgiy A |
author_sort | Prajapati, Ranjit K |
collection | PubMed |
description | Oxazinomycin is a C-nucleoside antibiotic that is produced by Streptomyces hygroscopicus and closely resembles uridine. Here, we show that the oxazinomycin triphosphate is a good substrate for bacterial and eukaryotic RNA polymerases (RNAPs) and that a single incorporated oxazinomycin is rapidly extended by the next nucleotide. However, the incorporation of several successive oxazinomycins or a single oxazinomycin in a certain sequence context arrested a fraction of the transcribing RNAP. The addition of Gre RNA cleavage factors eliminated the transcriptional arrest at a single oxazinomycin and shortened the nascent RNAs arrested at the polythymidine sequences suggesting that the transcriptional arrest was caused by backtracking of RNAP along the DNA template. We further demonstrate that the ubiquitous C-nucleoside pseudouridine is also a good substrate for RNA polymerases in a triphosphorylated form but does not inhibit transcription of the polythymidine sequences. Our results collectively suggest that oxazinomycin functions as a Trojan horse substrate and its inhibitory effect is attributable to the oxygen atom in the position corresponding to carbon five of the uracil ring. |
format | Online Article Text |
id | pubmed-6821320 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-68213202019-11-04 Oxazinomycin arrests RNA polymerase at the polythymidine sequences Prajapati, Ranjit K Rosenqvist, Petja Palmu, Kaisa Mäkinen, Janne J Malinen, Anssi M Virta, Pasi Metsä-Ketelä, Mikko Belogurov, Georgiy A Nucleic Acids Res Nucleic Acid Enzymes Oxazinomycin is a C-nucleoside antibiotic that is produced by Streptomyces hygroscopicus and closely resembles uridine. Here, we show that the oxazinomycin triphosphate is a good substrate for bacterial and eukaryotic RNA polymerases (RNAPs) and that a single incorporated oxazinomycin is rapidly extended by the next nucleotide. However, the incorporation of several successive oxazinomycins or a single oxazinomycin in a certain sequence context arrested a fraction of the transcribing RNAP. The addition of Gre RNA cleavage factors eliminated the transcriptional arrest at a single oxazinomycin and shortened the nascent RNAs arrested at the polythymidine sequences suggesting that the transcriptional arrest was caused by backtracking of RNAP along the DNA template. We further demonstrate that the ubiquitous C-nucleoside pseudouridine is also a good substrate for RNA polymerases in a triphosphorylated form but does not inhibit transcription of the polythymidine sequences. Our results collectively suggest that oxazinomycin functions as a Trojan horse substrate and its inhibitory effect is attributable to the oxygen atom in the position corresponding to carbon five of the uracil ring. Oxford University Press 2019-11-04 2019-09-09 /pmc/articles/PMC6821320/ /pubmed/31495891 http://dx.doi.org/10.1093/nar/gkz782 Text en © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Nucleic Acid Enzymes Prajapati, Ranjit K Rosenqvist, Petja Palmu, Kaisa Mäkinen, Janne J Malinen, Anssi M Virta, Pasi Metsä-Ketelä, Mikko Belogurov, Georgiy A Oxazinomycin arrests RNA polymerase at the polythymidine sequences |
title | Oxazinomycin arrests RNA polymerase at the polythymidine sequences |
title_full | Oxazinomycin arrests RNA polymerase at the polythymidine sequences |
title_fullStr | Oxazinomycin arrests RNA polymerase at the polythymidine sequences |
title_full_unstemmed | Oxazinomycin arrests RNA polymerase at the polythymidine sequences |
title_short | Oxazinomycin arrests RNA polymerase at the polythymidine sequences |
title_sort | oxazinomycin arrests rna polymerase at the polythymidine sequences |
topic | Nucleic Acid Enzymes |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6821320/ https://www.ncbi.nlm.nih.gov/pubmed/31495891 http://dx.doi.org/10.1093/nar/gkz782 |
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