This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Bangur, C. S. Articles by Ponticelli, A. S. Search for Related Content PubMed PubMed Citation Articles by Bangur, C. S. Articles by Ponticelli, A. S.

 Previous Article  |  Next Article 

Mol. Cell. Biol., 12 1997, 6784-6793, Vol 17, No. 12
Copyright © 1997, American Society for Microbiology

Mutational analysis of the D1/E1 core helices and the conserved N- terminal region of yeast transcription factor IIB (TFIIB): identification of an N-terminal mutant that stabilizes TATA-binding protein-TFIIB-DNA complexes

CS Bangur, TS Pardee and AS Ponticelli
Department of Biochemistry and Center for Advanced Molecular Biology and Immunology, School of Medicine and Biomedical Sciences, State University of New York at Buffalo, 14214-3000, USA.

The general transcription factor IIB (TFIIB) plays an essential role in transcription of protein-coding genes by RNA polymerase II. We have used site-directed mutagenesis to assess the role of conserved amino acids in several important regions of yeast TFIIB. These include residues in the highly conserved amino-terminal region and basic residues in the D1 and E1 core domain alpha-helices. Acidic substitutions of residues K190 (D1) and K201 (E1) resulted in growth impairments in vivo, reduced basal transcriptional activity in vitro, and an inability to form stable TFIIB-TATA-binding protein-DNA (DB) complexes. Significantly, these mutants retained the ability to respond to acidic activators in vivo and to the Gal4-VP16 activator in vitro, supporting the view that these basic residues play a role in basal transcription. In addition, 14 single-amino-acid substitutions were introduced in the conserved amino-terminal region. Three of these mutants, the L50D, R64E, and R78L mutants, displayed altered growth properties in vivo and were compromised for supporting transcription in vitro. The L50D mutant was impaired for RNA polymerase II interaction, while the R64E mutant exhibited altered transcription start site selection both in vitro and in vivo and, surprisingly, was more active than the wild type in the formation of stable DB complexes. These results support the view that the amino-terminal domain is involved in the direct interaction between yeast TFIIB and RNA polymerase II and suggest that this domain may interact with DNA and/or modulate the formation of a DB complex.

This article has been cited by other articles:

• Khaperskyy, D. A., Ammerman, M. L., Majovski, R. C., Ponticelli, A. S. (2008). Functions of Saccharomyces cerevisiae TFIIF during Transcription Start Site Utilization. Mol. Cell. Biol. 28: 3757-3766 [Abstract] [Full Text]  
• Sevilimedu, A., Shi, H., Lis, J. T. (2008). TFIIB aptamers inhibit transcription by perturbing PIC formation at distinct stages. Nucleic Acids Res 36: 3118-3127 [Abstract] [Full Text]  
• Kasahara, K., Ki, S., Aoyama, K., Takahashi, H., Kokubo, T. (2008). Saccharomyces cerevisiae HMO1 interacts with TFIID and participates in start site selection by RNA polymerase II. Nucleic Acids Res 36: 1343-1357 [Abstract] [Full Text]  
• Majovski, R. C., Khaperskyy, D. A., Ghazy, M. A., Ponticelli, A. S. (2005). A Functional Role for the Switch 2 Region of Yeast RNA Polymerase II in Transcription Start Site Utilization and Abortive Initiation. J. Biol. Chem. 280: 34917-34923 [Abstract] [Full Text]  
• Chen, B.-S., Hampsey, M. (2004). Functional Interaction between TFIIB and the Rpb2 Subunit of RNA Polymerase II: Implications for the Mechanism of Transcription Initiation. Mol. Cell. Biol. 24: 3983-3991 [Abstract] [Full Text]  
• Tubon, T. C., Tansey, W. P., Herr, W. (2004). A Nonconserved Surface of the TFIIB Zinc Ribbon Domain Plays a Direct Role in RNA Polymerase II Recruitment. Mol. Cell. Biol. 24: 2863-2874 [Abstract] [Full Text]  
• Glossop, J. A., Dafforn, T. R., Roberts, S. G. E. (2004). A conformational change in TFIIB is required for activator-mediated assembly of the preinitiation complex. Nucleic Acids Res 32: 1829-1835 [Abstract] [Full Text]  
• Bushnell, D. A., Westover, K. D., Davis, R. E., Kornberg, R. D. (2004). Structural Basis of Transcription: An RNA Polymerase II-TFIIB Cocrystal at 4.5 Angstroms. Science 303: 983-988 [Abstract] [Full Text]  
• Renfrow, M. B., Naryshkin, N., Lewis, L. M., Chen, H.-T., Ebright, R. H., Scott, R. A. (2004). Transcription Factor B Contacts Promoter DNA Near the Transcription Start Site of the Archaeal Transcription Initiation Complex. J. Biol. Chem. 279: 2825-2831 [Abstract] [Full Text]  
• Ziegler, L. M., Khaperskyy, D. A., Ammerman, M. L., Ponticelli, A. S. (2003). Yeast RNA Polymerase II Lacking the Rpb9 Subunit Is Impaired for Interaction with Transcription Factor IIF. J. Biol. Chem. 278: 48950-48956 [Abstract] [Full Text]  
• Fairley, J. A., Evans, R., Hawkes, N. A., Roberts, S. G. E. (2002). Core Promoter-Dependent TFIIB Conformation and a Role for TFIIB Conformation in Transcription Start Site Selection. Mol. Cell. Biol. 22: 6697-6705 [Abstract] [Full Text]  
• Zhang, D.-Y., Carson, D. J., Ma, J. (2002). The role of TFIIB-RNA polymerase II interaction in start site selection in yeast cells. Nucleic Acids Res 30: 3078-3085 [Abstract] [Full Text]  
• Faitar, S. L., Brodie, S. A., Ponticelli, A. S. (2001). Promoter-Specific Shifts in Transcription Initiation Conferred by Yeast TFIIB Mutations Are Determined by the Sequence in the Immediate Vicinity of the Start Sites. Mol. Cell. Biol. 21: 4427-4440 [Abstract] [Full Text]  
• Kobor, M. S., Simon, L. D., Omichinski, J., Zhong, G., Archambault, J., Greenblatt, J. (2000). A Motif Shared by TFIIF and TFIIB Mediates Their Interaction with the RNA Polymerase II Carboxy-Terminal Domain Phosphatase Fcp1p in Saccharomyces cerevisiae. Mol. Cell. Biol. 20: 7438-7449 [Abstract] [Full Text]  
• Zhang, D.-Y., Dorsey, M. J., Voth, W. P., Carson, D. J., Zeng, X., Stillman, D. J., Ma, J. (2000). Intramolecular interaction of yeast TFIIB in transcription control. Nucleic Acids Res 28: 1913-1920 [Abstract] [Full Text]  
• Bell, S. D., Jackson, S. P. (2000). The Role of Transcription Factor B in Transcription Initiation and Promoter Clearance in the Archaeon Sulfolobus acidocaldarius. J. Biol. Chem. 275: 12934-12940 [Abstract] [Full Text]  
• Hahn, S., Roberts, S. (2000). The zinc ribbon domains of the general transcription factors TFIIB and Brf: conserved functional surfaces but different roles in transcription initiation. Genes Dev. 14: 719-730 [Abstract] [Full Text]  
• Serpe, M., Joshi, A., Kosman, D. J. (1999). Structure-Function Analysis of the Protein-binding Domains of Mac1p, a Copper-dependent Transcriptional Activator of Copper Uptake in Saccharomyces cerevisiae. J. Biol. Chem. 274: 29211-29219 [Abstract] [Full Text]  
• Wu, W.-H., Pinto, I., Chen, B.-S., Hampsey, M. (1999). Mutational Analysis of Yeast TFIIB: A Functional Relationship Between Ssu72 and Sub1/Tsp1 Defined by Allele-Specific Interactions With TFIIB. Genetics 153: 643-652 [Abstract] [Full Text]  
• Cho, E.-J., Buratowski, S. (1999). Evidence That Transcription Factor IIB Is Required for a Post-assembly Step in Transcription Initiation. J. Biol. Chem. 274: 25807-25813 [Abstract] [Full Text]  
• Bangur, C. S., Faitar, S. L., Folster, J. P., Ponticelli, A. S. (1999). An Interaction between the N-terminal Region and the Core Domain of Yeast TFIIB Promotes the Formation of TATA-binding Protein-TFIIB-DNA Complexes. J. Biol. Chem. 274: 23203-23209 [Abstract] [Full Text]  
• Hawkes, N. A., Roberts, S. G. E. (1999). The Role of Human TFIIB in Transcription Start Site Selection in Vitro and in Vivo. J. Biol. Chem. 274: 14337-14343 [Abstract] [Full Text]  
• Pardee, T. S., Bangur, C. S., Ponticelli, A. S. (1998). The N-terminal Region of Yeast TFIIB Contains Two Adjacent Functional Domains Involved in Stable RNA Polymerase II Binding and Transcription Start Site Selection. J. Biol. Chem. 273: 17859-17864 [Abstract] [Full Text]  
• Hampsey, M. (1998). Molecular Genetics of the RNA Polymerase II General Transcriptional Machinery. Microbiol. Mol. Biol. Rev. 62: 465-503 [Abstract] [Full Text]