PuSH - Publication Server of Helmholtz Zentrum München

Export: TXT Text RIS Endnote (RIS) BIB BibTeX
Ghamari, A.* ; van de Corput, M.P.C.* ; Thongjuea, S.* ; van Cappellen, W.A.* ; van Ijcken, W.* ; van Haren, J.* ; Soler, E.* ; Eick, D. ; Lenhard, B.* ; Grosveld, F.G.*

In vivo live imaging of RNA polymerase II transcription factories in primary cells.

Genes Dev. 27, 767-777 (2013)
Publ. Version/Full Text Volltext DOI PMC
Closed
Transcription steps are marked by different modifications of the C-terminal domain of RNA polymerase II (RNAPII). Phosphorylation of Ser5 and Ser7 by cyclin-dependent kinase 7 (CDK7) as part of TFIIH marks initiation, whereas phosphorylation of Ser2 by CDK9 marks elongation. These processes are thought to take place in localized transcription foci in the nucleus, known as "transcription factories,'' but it has been argued that the observed clusters/foci are mere fixation or labeling artifacts. We show that transcription factories exist in living cells as distinct foci by live-imaging fluorescently labeled CDK9, a kinase known to associate with active RNAPII. These foci were observed in different cell types derived from CDK9-mCherry knock-in mice. We show that these foci are very stable while highly dynamic in exchanging CDK9. Chromatin immunoprecipitation (ChIP) coupled with deep sequencing (ChIP-seq) data show that the genome-wide binding sites of CDK9 and initiating RNAPII overlap on transcribed genes. Immunostaining shows that CDK9-mCherry foci colocalize with RNAPII-Ser5P, much less with RNAPII-Ser2P, and not with CDK12 (a kinase reported to be involved in the Ser2 phosphorylation) or with splicing factor SC35. In conclusion, transcription factories exist in living cells, and initiation and elongation of transcripts takes place in different nuclear compartments.
Impact Factor
Scopus SNIP
Web of Science
Times Cited
Scopus
Cited By
Altmetric
12.444
2.415
92
97
Tags
Annotations
Special Publikation
Hide on homepage

Edit extra information
Edit own tags
Private
Edit own annotation
Private
Hide on publication lists
on hompage
Mark as special
publikation
Publication type Article: Journal article
Document type Scientific Article
Keywords Transcription Factory ; Rna Polymerase Ii ; Live Imaging; P-tefb ; Gene-expression ; Human Nuclei ; 7sk Snrna ; Elongation ; Kinase ; Organization ; Phosphorylation ; Flavopiridol ; Association
Language english
Publication Year 2013
HGF-reported in Year 2013
ISSN (print) / ISBN 0890-9369
e-ISSN 1549-5477
Journal Genes and Development
Quellenangaben Volume: 27, Issue: 7, Pages: 767-777 Article Number: , Supplement: ,
Publisher Cold Spring Harbor Laboratory Press
Reviewing status Peer reviewed
Institute(s) Institute of Clinical Molecular Biology and Tumor Genetics (K.MOLBI)
POF-Topic(s) 30504 - Mechanisms of Genetic and Environmental Influences on Health and Disease
Research field(s) Immune Response and Infection
PSP Element(s) G-501490-001
PubMed ID 23592796
DOI 10.1101/gad.216200.113
WOS ID WOS:000317578000006
Scopus ID 84876276025
Erfassungsdatum 2013-05-24
[➜Report correction]