Article
- The EMBO Journal (2009) 28, 854 - 865
- doi:10.1038/emboj.2009.33
Published online: 12 February 2009
Subject Category:
FACT facilitates chromatin transcription by RNA polymerases I and IIIEMBO Open
Joanna L Birch1,a, Bertrand C-M Tan2,a, Kostya I Panov1,b, Tatiana B Panova1,c, Jens S Andersen3, Tom A Owen-Hughes1, Jackie Russell1, Sheng-Chung Lee4,5 and Joost C B M Zomerdijk1
- Wellcome Trust Centre for Gene Regulation and Expression, College of Life Sciences, University of Dundee, Dundee, UK
- Department of Life Sciences, Chang Gung University, Kwei-Shan Tao-Yuan, Taiwan
- Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark
- Institute of Molecular Medicine, College of Medicine, National Taiwan University, Taipei, Taiwan
- Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan
Correspondence to:
Joost C B M Zomerdijk, Wellcome Trust Centre for Gene Regulation and Expression, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK. Tel.: +44 1382 384 242; Fax: +44 1382 388 072; E-mail: j.zomerdijk@dundee.ac.uk
aThese authors contributed equally to this work
bPresent address: The School of Biological Sciences, Queen's University Belfast, Belfast BT7 1NN, UK
cPresent address: Dundee Cell Products Ltd, Dundee Technopole, Dundee DD1 5JJ, UK
Received 13 August 2008; Accepted 21 January 2009
Abstract
Efficient transcription elongation from a chromatin template requires RNA polymerases (Pols) to negotiate nucleosomes. Our biochemical analyses demonstrate that RNA Pol I can transcribe through nucleosome templates and that this requires structural rearrangement of the nucleosomal core particle. The subunits of the histone chaperone FACT (facilitates chromatin transcription), SSRP1 and Spt16, co-purify and co-immunoprecipitate with mammalian Pol I complexes. In cells, SSRP1 is detectable at the rRNA gene repeats. Crucially, siRNA-mediated repression of FACT subunit expression in cells results in a significant reduction in 47S pre-rRNA levels, whereas synthesis of the first 40 nt of the rRNA is not affected, implying that FACT is important for Pol I transcription elongation through chromatin. FACT also associates with RNA Pol III complexes, is present at the chromatin of genes transcribed by Pol III and facilitates their transcription in cells. Our findings indicate that, beyond the established role in Pol II transcription, FACT has physiological functions in chromatin transcription by all three nuclear RNA Pols. Our data also imply that local chromatin dynamics influence transcription of the active rRNA genes by Pol I and of Pol III-transcribed genes.
Keywords:
- rDNA chromatin,
- ribosomal RNA,
- transcription elongation,
- Spt16,
- SSRP1
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