![]()
|
|
||||||||
J. Biol. Chem., Vol. 281, Issue 17, 11685-11692, April 28, 2006
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
From the Service de Biochimie et de Génétique Moléculaire, Bâtiment 144, CEA/Saclay, F-91191 Gif-sur-Yvette Cedex, France
Transcription factor TFIIIC is a multisubunit complex required for promoter recognition and transcriptional activation of class III genes. We describe here the reconstitution of complete recombinant yeast TFIIIC and the molecular characterization of its two DNA-binding domains,
A and
B, using the baculovirus expression system. The B block-binding module, r
B, was reconstituted with r
138, r
91, and r
60 subunits. r
131, r
95, and r
55 formed also a stable complex, r
A, that displayed nonspecific DNA binding activity. Recombinant rTFIIIC was functionally equivalent to purified yeast TFIIIC, suggesting that the six recombinant subunits are necessary and sufficient to reconstitute a transcriptionally active TFIIIC complex. The formation and the properties of rTFIIIC-DNA complexes were affected by dephosphorylation treatments. The combination of complete recombinant rTFIIIC and rTFIIIB directed a low level of basal transcription, much weaker than with the crude B'' fraction, suggesting the existence of auxiliary factors that could modulate the yeast RNA polymerase III transcription system.
Received for publication, January 5, 2006 , and in revised form, February 23, 2006.
* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
1 Present address: CMU/Département de Microbiologie et de Médecine Moléculaire,Genève, Suisse.
2 Present address: Laboratory of Molecular Pharmacology, Center for Cancer Research, National Cancer Institute, National Institute of Health, Bethesda, MD 20892-4255.
3 To whom correspondence should be addressed. Tel.: 33-1-69083796; Fax: 33-1-69084712; E-mail: joel.acker{at}cea.fr.
![]()
CiteULike
Complore
Connotea
Del.icio.us
Digg
Reddit
Technorati What's this?
This article has been cited by other articles:
![]() |
Y. Ghavi-Helm, M. Michaut, J. Acker, J.-C. Aude, P. Thuriaux, M. Werner, and J. Soutourina Genome-wide location analysis reveals a role of TFIIS in RNA polymerase III transcription Genes & Dev., July 15, 2008; 22(14): 1934 - 1947. [Abstract] [Full Text] [PDF] |
||||
![]() |
H. Dumay-Odelot, C. Marck, S. Durrieu-Gaillard, O. Lefebvre, S. Jourdain, M. Prochazkova, A. Pflieger, and M. Teichmann Identification, Molecular Cloning, and Characterization of the Sixth Subunit of Human Transcription Factor TFIIIC J. Biol. Chem., June 8, 2007; 282(23): 17179 - 17189. [Abstract] [Full Text] [PDF] |
||||
![]() |
E. Guffanti, R. Ferrari, M. Preti, M. Forloni, O. Harismendy, O. Lefebvre, and G. Dieci A Minimal Promoter for TFIIIC-dependent in Vitro Transcription of snoRNA and tRNA Genes by RNA Polymerase III J. Biol. Chem., August 18, 2006; 281(33): 23945 - 23957. [Abstract] [Full Text] [PDF] |
||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |