Transition from initiation to promoter proximal pausing requires the CTD of RNA polymerase II

The C-terminal domain (CTD) of mammalian RNA polymerase II consists of 52 repeats of the consensus hepta-peptide YSPTSPS, and links transcription to the processing of pre-mRNA. Although Pol II with a CTD shortened to five repeats (Pol II Δ5) is transcriptionally inactive on chromatin templates, it i...

Full description

Bibliographic Details
Main Authors:	Lux, C., Albiez, H., Chapman, R. D., Heidinger, M., Meininghaus, M., Brack-Werner, R., Lang, A., Ziegler, M., Cremer, T., Eick, D.
Format:	Online
Language:	English
Published:	Oxford University Press 2005
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1214543/

id	pubmed-1214543
recordtype	oai_dc
spelling	pubmed-12145432005-09-15 Transition from initiation to promoter proximal pausing requires the CTD of RNA polymerase II Lux, C. Albiez, H. Chapman, R. D. Heidinger, M. Meininghaus, M. Brack-Werner, R. Lang, A. Ziegler, M. Cremer, T. Eick, D. Molecular Biology The C-terminal domain (CTD) of mammalian RNA polymerase II consists of 52 repeats of the consensus hepta-peptide YSPTSPS, and links transcription to the processing of pre-mRNA. Although Pol II with a CTD shortened to five repeats (Pol II Δ5) is transcriptionally inactive on chromatin templates, it is not clear whether CTD is required for promoter recognition in vivo. Here, we demonstrate that in the context of chromatin, Pol II Δ5 can bind to the c-myc promoter with the same efficiency as wild type Pol II. However, Pol II Δ5 does not form a stable initiation complex, and does not transcribe promoter proximal sequences. Fluorescence recovery after photobleaching (FRAP) experiments with cells expressing enhanced green fluorescent protein (EGFP)-tagged Δ5 or wildtype Pol II revealed a single, highly mobile Pol II Δ5 fraction whereas wildtype Pol II yielded less mobile fractions. These data suggest that CTD is not required for promoter recognition, but rather for subsequent formation of a stable initiation complex and isomerization to an elongation competent complex. Oxford University Press 2005 2005-09-12 /pmc/articles/PMC1214543/ /pubmed/16157863 http://dx.doi.org/10.1093/nar/gki802 Text en © The Author 2005. Published by Oxford University Press. All rights reserved
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Lux, C. Albiez, H. Chapman, R. D. Heidinger, M. Meininghaus, M. Brack-Werner, R. Lang, A. Ziegler, M. Cremer, T. Eick, D.
spellingShingle	Lux, C. Albiez, H. Chapman, R. D. Heidinger, M. Meininghaus, M. Brack-Werner, R. Lang, A. Ziegler, M. Cremer, T. Eick, D. Transition from initiation to promoter proximal pausing requires the CTD of RNA polymerase II
author_facet	Lux, C. Albiez, H. Chapman, R. D. Heidinger, M. Meininghaus, M. Brack-Werner, R. Lang, A. Ziegler, M. Cremer, T. Eick, D.
author_sort	Lux, C.
title	Transition from initiation to promoter proximal pausing requires the CTD of RNA polymerase II
title_short	Transition from initiation to promoter proximal pausing requires the CTD of RNA polymerase II
title_full	Transition from initiation to promoter proximal pausing requires the CTD of RNA polymerase II
title_fullStr	Transition from initiation to promoter proximal pausing requires the CTD of RNA polymerase II
title_full_unstemmed	Transition from initiation to promoter proximal pausing requires the CTD of RNA polymerase II
title_sort	transition from initiation to promoter proximal pausing requires the ctd of rna polymerase ii
description	The C-terminal domain (CTD) of mammalian RNA polymerase II consists of 52 repeats of the consensus hepta-peptide YSPTSPS, and links transcription to the processing of pre-mRNA. Although Pol II with a CTD shortened to five repeats (Pol II Δ5) is transcriptionally inactive on chromatin templates, it is not clear whether CTD is required for promoter recognition in vivo. Here, we demonstrate that in the context of chromatin, Pol II Δ5 can bind to the c-myc promoter with the same efficiency as wild type Pol II. However, Pol II Δ5 does not form a stable initiation complex, and does not transcribe promoter proximal sequences. Fluorescence recovery after photobleaching (FRAP) experiments with cells expressing enhanced green fluorescent protein (EGFP)-tagged Δ5 or wildtype Pol II revealed a single, highly mobile Pol II Δ5 fraction whereas wildtype Pol II yielded less mobile fractions. These data suggest that CTD is not required for promoter recognition, but rather for subsequent formation of a stable initiation complex and isomerization to an elongation competent complex.
publisher	Oxford University Press
publishDate	2005
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1214543/
_version_	1611376730742718464

Transition from initiation to promoter proximal pausing requires the CTD of RNA polymerase II

Similar Items