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Molecular and Cellular Biology, November 2004, p. 10072-10082, Vol. 24, No. 22
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.22.10072-10082.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

NC2{alpha} Interacts with BTAF1 and Stimulates Its ATP-Dependent Association with TATA-Binding Protein

Marcin P. Klejman,1 Lloyd A. Pereira,1,{dagger} Hester J. T. van Zeeburg,1,{ddagger} Siv Gilfillan,2 Michael Meisterernst,2 and H. T. Marc Timmers1*

Department of Physiological Chemistry, Division of Biomedical Genetics, University Medical Center Utrecht, Utrecht, The Netherlands,1 National Research Center for Environment and Health-GSF, Institute of Molecular Immunology, Gene Expression, Munich, Germany2

Received 22 July 2004/ Accepted 21 August 2004

Transcriptional activity of the TATA-binding protein (TBP) is controlled by a variety of proteins. The BTAF1 protein (formerly known as TAFII170/TAF-172 and the human ortholog of Saccharomyces cerevisiae Mot1p) and the NC2 complex composed of NC2{alpha} (DRAP1) and NC2ß (Dr1) are able to bind to TBP directly and regulate RNA polymerase II transcription both positively and negatively. Here, we present evidence that the NC2{alpha} subunit interacts with BTAF1. In contrast, the NC2ß subunit is not able to associate with BTAF1 and seems to interfere with the BTAF1-TBP interaction. Addition of NC2{alpha} or the NC2 complex can stimulate the ability of BTAF1 to interact with TBP. This function is dependent on the presence of ATP in cell extracts but does not involve the ATPase activity of BTAF1 nor phosphorylation of NC2{alpha}. Together, our results constitute the first evidence of the physical cooperation between BTAF1 and NC2{alpha} in TBP regulation and provide a framework to understand transcription functions of NC2{alpha} and NC2ß in vivo.

* Corresponding author. Mailing address: Department of Physiological Chemistry, Division of Biomedical Genetics, University Medical Center Utrecht, Universiteitsweg 100, 3584 CG Utrecht, The Netherlands. Phone: 31 30 253 8981. Fax: 31 30 253 9035. E-mail: h.t.m.timmers{at}med.uu.nl.

{dagger} Present address: Centre for Early Human Development, Monash Institute for Reproduction and Development, Clayton, Victoria 3168, Australia.

{ddagger} Present address: Section of Tumor Biology, Department of Otolaryngology/Head and Neck Surgery, Vrije Universiteit Medical Center, 1007 MB Amsterdam, The Netherlands.

Molecular and Cellular Biology, November 2004, p. 10072-10082, Vol. 24, No. 22
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.22.10072-10082.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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