Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Prajapati, Ranjit K, Rosenqvist, Petja, Palmu, Kaisa, Mäkinen, Janne J, Malinen, Anssi M, Virta, Pasi, Metsä-Ketelä, Mikko, Belogurov, Georgiy A
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2019
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
_version_ 1783464122564739072
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
work_keys_str_mv AT prajapatiranjitk oxazinomycinarrestsrnapolymeraseatthepolythymidinesequences
AT rosenqvistpetja oxazinomycinarrestsrnapolymeraseatthepolythymidinesequences
AT palmukaisa oxazinomycinarrestsrnapolymeraseatthepolythymidinesequences
AT makinenjannej oxazinomycinarrestsrnapolymeraseatthepolythymidinesequences
AT malinenanssim oxazinomycinarrestsrnapolymeraseatthepolythymidinesequences
AT virtapasi oxazinomycinarrestsrnapolymeraseatthepolythymidinesequences
AT metsaketelamikko oxazinomycinarrestsrnapolymeraseatthepolythymidinesequences
AT belogurovgeorgiya oxazinomycinarrestsrnapolymeraseatthepolythymidinesequences