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RNA Polymerase II Activity of Type 3 Pol III Promoters

In eukaryotes, three RNA polymerases (Pol I, II, and III) are responsible for the transcription of distinct subsets of genes. Gene-external type 3 Pol III promoters use defined transcription start and termination sites, and they are, therefore, widely used for small RNA expression, including short h...

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Detalles Bibliográficos
Autores principales: Gao, Zongliang, Herrera-Carrillo, Elena, Berkhout, Ben
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Gene & Cell Therapy 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6023835/
https://www.ncbi.nlm.nih.gov/pubmed/30195753
http://dx.doi.org/10.1016/j.omtn.2018.05.001
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author Gao, Zongliang
Herrera-Carrillo, Elena
Berkhout, Ben
author_facet Gao, Zongliang
Herrera-Carrillo, Elena
Berkhout, Ben
author_sort Gao, Zongliang
collection PubMed
description In eukaryotes, three RNA polymerases (Pol I, II, and III) are responsible for the transcription of distinct subsets of genes. Gene-external type 3 Pol III promoters use defined transcription start and termination sites, and they are, therefore, widely used for small RNA expression, including short hairpin RNAs in RNAi applications and guide RNAs in CRISPR-Cas systems. We report that all three commonly used human Pol III promoters (7SK, U6, and H1) mediate luciferase reporter gene expression, which indicates Pol II activity, but to a different extent (H1 ≫ U6 > 7SK). We demonstrate that these promoters can recruit Pol II for transcribing extended messenger transcripts. Intriguingly, selective inhibition of Pol II stimulates the Pol III activity and vice versa, suggesting that two polymerase complexes compete for promoter usage. Pol II initiates transcription at the regular Pol III start site on the 7SK and U6 promoters, but Pol II transcription on the most active H1 promoter starts 8 nt upstream of the Pol III start site. This study provides functional evidence for the close relationship of Pol II and Pol III transcription. These mechanistic insights are important for optimal use of Pol III promoters, and they offer additional flexibility for biotechnology applications of these genetic elements.
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spelling pubmed-60238352018-06-29 RNA Polymerase II Activity of Type 3 Pol III Promoters Gao, Zongliang Herrera-Carrillo, Elena Berkhout, Ben Mol Ther Nucleic Acids Article In eukaryotes, three RNA polymerases (Pol I, II, and III) are responsible for the transcription of distinct subsets of genes. Gene-external type 3 Pol III promoters use defined transcription start and termination sites, and they are, therefore, widely used for small RNA expression, including short hairpin RNAs in RNAi applications and guide RNAs in CRISPR-Cas systems. We report that all three commonly used human Pol III promoters (7SK, U6, and H1) mediate luciferase reporter gene expression, which indicates Pol II activity, but to a different extent (H1 ≫ U6 > 7SK). We demonstrate that these promoters can recruit Pol II for transcribing extended messenger transcripts. Intriguingly, selective inhibition of Pol II stimulates the Pol III activity and vice versa, suggesting that two polymerase complexes compete for promoter usage. Pol II initiates transcription at the regular Pol III start site on the 7SK and U6 promoters, but Pol II transcription on the most active H1 promoter starts 8 nt upstream of the Pol III start site. This study provides functional evidence for the close relationship of Pol II and Pol III transcription. These mechanistic insights are important for optimal use of Pol III promoters, and they offer additional flexibility for biotechnology applications of these genetic elements. American Society of Gene & Cell Therapy 2018-05-08 /pmc/articles/PMC6023835/ /pubmed/30195753 http://dx.doi.org/10.1016/j.omtn.2018.05.001 Text en © 2018 The Author(s) http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Gao, Zongliang
Herrera-Carrillo, Elena
Berkhout, Ben
RNA Polymerase II Activity of Type 3 Pol III Promoters
title RNA Polymerase II Activity of Type 3 Pol III Promoters
title_full RNA Polymerase II Activity of Type 3 Pol III Promoters
title_fullStr RNA Polymerase II Activity of Type 3 Pol III Promoters
title_full_unstemmed RNA Polymerase II Activity of Type 3 Pol III Promoters
title_short RNA Polymerase II Activity of Type 3 Pol III Promoters
title_sort rna polymerase ii activity of type 3 pol iii promoters
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6023835/
https://www.ncbi.nlm.nih.gov/pubmed/30195753
http://dx.doi.org/10.1016/j.omtn.2018.05.001
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