The in vivo kinetics of RNA polymerase II elongation during co-transcriptional splicing.

RNA processing events that take place on the transcribed pre-mRNA include capping, splicing, editing, 3' processing, and polyadenylation. Most of these processes occur co-transcriptionally while the RNA polymerase II (Pol II) enzyme is engaged in transcriptional elongation. How Pol II elongatio...

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Autores principales:	Yehuda Brody, Noa Neufeld, Nicole Bieberstein, Sebastien Z Causse, Eva-Maria Böhnlein, Karla M Neugebauer, Xavier Darzacq, Yaron Shav-Tal
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/a1d350b1dd2444d7b078f4630d428359
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spelling	oai:doaj.org-article:a1d350b1dd2444d7b078f4630d4283592021-11-18T05:36:22ZThe in vivo kinetics of RNA polymerase II elongation during co-transcriptional splicing.1544-91731545-788510.1371/journal.pbio.1000573https://doaj.org/article/a1d350b1dd2444d7b078f4630d4283592011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21264352/pdf/?tool=EBIhttps://doaj.org/toc/1544-9173https://doaj.org/toc/1545-7885RNA processing events that take place on the transcribed pre-mRNA include capping, splicing, editing, 3' processing, and polyadenylation. Most of these processes occur co-transcriptionally while the RNA polymerase II (Pol II) enzyme is engaged in transcriptional elongation. How Pol II elongation rates are influenced by splicing is not well understood. We generated a family of inducible gene constructs containing increasing numbers of introns and exons, which were stably integrated in human cells to serve as actively transcribing gene loci. By monitoring the association of the transcription and splicing machineries on these genes in vivo, we showed that only U1 snRNP localized to the intronless gene, consistent with a splicing-independent role for U1 snRNP in transcription. In contrast, all snRNPs accumulated on intron-containing genes, and increasing the number of introns increased the amount of spliceosome components recruited. This indicates that nascent RNA can assemble multiple spliceosomes simultaneously. Kinetic measurements of Pol II elongation in vivo, Pol II ChIP, as well as use of Spliceostatin and Meayamycin splicing inhibitors showed that polymerase elongation rates were uncoupled from ongoing splicing. This study shows that transcription elongation kinetics proceed independently of splicing at the model genes studied here. Surprisingly, retention of polyadenylated mRNA was detected at the transcription site after transcription termination. This suggests that the polymerase is released from chromatin prior to the completion of splicing, and the pre-mRNA is post-transcriptionally processed while still tethered to chromatin near the gene end.Yehuda BrodyNoa NeufeldNicole BiebersteinSebastien Z CausseEva-Maria BöhnleinKarla M NeugebauerXavier DarzacqYaron Shav-TalPublic Library of Science (PLoS)articleBiology (General)QH301-705.5ENPLoS Biology, Vol 9, Iss 1, p e1000573 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Biology (General) QH301-705.5
spellingShingle	Biology (General) QH301-705.5 Yehuda Brody Noa Neufeld Nicole Bieberstein Sebastien Z Causse Eva-Maria Böhnlein Karla M Neugebauer Xavier Darzacq Yaron Shav-Tal The in vivo kinetics of RNA polymerase II elongation during co-transcriptional splicing.
description	RNA processing events that take place on the transcribed pre-mRNA include capping, splicing, editing, 3' processing, and polyadenylation. Most of these processes occur co-transcriptionally while the RNA polymerase II (Pol II) enzyme is engaged in transcriptional elongation. How Pol II elongation rates are influenced by splicing is not well understood. We generated a family of inducible gene constructs containing increasing numbers of introns and exons, which were stably integrated in human cells to serve as actively transcribing gene loci. By monitoring the association of the transcription and splicing machineries on these genes in vivo, we showed that only U1 snRNP localized to the intronless gene, consistent with a splicing-independent role for U1 snRNP in transcription. In contrast, all snRNPs accumulated on intron-containing genes, and increasing the number of introns increased the amount of spliceosome components recruited. This indicates that nascent RNA can assemble multiple spliceosomes simultaneously. Kinetic measurements of Pol II elongation in vivo, Pol II ChIP, as well as use of Spliceostatin and Meayamycin splicing inhibitors showed that polymerase elongation rates were uncoupled from ongoing splicing. This study shows that transcription elongation kinetics proceed independently of splicing at the model genes studied here. Surprisingly, retention of polyadenylated mRNA was detected at the transcription site after transcription termination. This suggests that the polymerase is released from chromatin prior to the completion of splicing, and the pre-mRNA is post-transcriptionally processed while still tethered to chromatin near the gene end.
format	article
author	Yehuda Brody Noa Neufeld Nicole Bieberstein Sebastien Z Causse Eva-Maria Böhnlein Karla M Neugebauer Xavier Darzacq Yaron Shav-Tal
author_facet	Yehuda Brody Noa Neufeld Nicole Bieberstein Sebastien Z Causse Eva-Maria Böhnlein Karla M Neugebauer Xavier Darzacq Yaron Shav-Tal
author_sort	Yehuda Brody
title	The in vivo kinetics of RNA polymerase II elongation during co-transcriptional splicing.
title_short	The in vivo kinetics of RNA polymerase II elongation during co-transcriptional splicing.
title_full	The in vivo kinetics of RNA polymerase II elongation during co-transcriptional splicing.
title_fullStr	The in vivo kinetics of RNA polymerase II elongation during co-transcriptional splicing.
title_full_unstemmed	The in vivo kinetics of RNA polymerase II elongation during co-transcriptional splicing.
title_sort	in vivo kinetics of rna polymerase ii elongation during co-transcriptional splicing.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/a1d350b1dd2444d7b078f4630d428359
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The in vivo kinetics of RNA polymerase II elongation during co-transcriptional splicing.

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