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Research Papers
Wellcome/CRC Institute, Cambridge University, UK.
Abstract
In Saccharomyces cerevisiae, two components of the RNA polymerase III (Pol III) general transcription factor TFIIIB are the TATA-binding protein (TBP) and the B-related factor (BRF), so called because its amino-terminal half is homologous to the Pol II transcription factor IIB (TFIIB). We have cloned BRF genes from the yeasts Kluyveromyces lactis and Candida albicans. Despite the large evolutionary distance between these species and S. cerevisiae, the BRF proteins are conserved highly. Although the homology is most pronounced in the amino-terminal half, conserved regions also exist in the carboxy-terminal half that is unique to BRF. By assaying for interactions between BRF and other Pol III transcription factors, we show that it is able to bind to the 135-kD subunit of TFIIIC and also to TBP. Surprisingly, in addition to binding the TFIIB-homologous amino-terminal portion of BRF, TBP also interacts strongly with the carboxy-terminal half. Deleting two conserved regions in the BRF carboxy-terminal region abrogates this interaction. Furthermore, TBP mutations that selectively inhibit Pol III transcription in vivo impair interactions between TBP and the BRF carboxy-terminal domain. Finally, we demonstrate that BRF but not TFIIB binds the Pol III subunit C34 and we define a region of C34 necessary for this interaction. These observations provide insights into the roles performed by BRF in Pol III transcription complex assembly.
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M. D. Librizzi, R. D. Moir, M. Brenowitz, and I. M. Willis Expression and Purification of the RNA Polymerase III Transcription Specificity Factor IIIB70 from Saccharomyces cerevisiae and Its Cooperative Binding with TATA-binding Protein J. Biol. Chem., December 20, 1996; 271(51): 32695 - 32701. [Abstract] [Full Text] [PDF]
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R. D. Moir, K. V. Puglia, and I. M. Willis Interactions between the Tetratricopeptide Repeat-containing Transcription Factor TFIIIC131 and Its Ligand, TFIIIB70. EVIDENCE FOR A CONFORMATIONAL CHANGE IN THE COMPLEX J. Biol. Chem., August 18, 2000; 275(34): 26591 - 26598. [Abstract] [Full Text] [PDF]
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T. E. Cloutier, M. D. Librizzi, A. K. M. M. Mollah, M. Brenowitz, and I. M. Willis Kinetic trapping of DNA by transcription factor IIIB PNAS, August 14, 2001; 98(17): 9581 - 9586. [Abstract] [Full Text] [PDF]
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