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 Lee, M. Articles by Struhl, K. Search for Related Content PubMed PubMed Citation Articles by Lee, M. Articles by Struhl, K.

 Previous Article  |  Next Article 

Mol. Cell. Biol., Oct 1995, 5461-5469, Vol 15, No. 10
Copyright © 1995, American Society for Microbiology

Mutations on the DNA-binding surface of TATA-binding protein can specifically impair the response to acidic activators in vivo

M Lee and K Struhl
Department Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.

The TATA-binding protein (TBP) contains a concave surface that interacts specifically with TATA promoter elements and a convex surface that mediates protein-protein interactions with general and gene- specific transcription factors. Biochemical experiments suggest that interactions between activator proteins and TBP are important in stimulating transcription by the RNA polymerase II machinery. To gain insight into the role of TBP in mediating transcriptional activation in vivo, we implemented a genetic strategy in Saccharomyces cerevisiae that involved the use of a TBP derivative with altered specificity for TATA elements. By genetically screening a set of TBP mutant libraries that were biased to the convex surface that mediates protein-protein interactions, we identified TBP derivatives that are impaired in the response to three acidic activators (Gcn4, Gal4, and Ace1) but appear normal for constitutive polymerase II transcription. A genetic complementation assay indicates that the activation-defective phenotypes reflect specific functional properties of the TBP derivatives rather than an indirect effect on transcription. Surprisingly, three of the four activation-defective mutants affect residues that directly contact DNA. Moreover, all four mutants are defective for TATA element binding, but they interact normally with an acidic activation domain and TFIIB. In addition, we show that a subset of TBP derivatives with mutations on the DNA-binding surface of TBP are also compromised in their responses to acidic activators in vivo. These observations suggest that interactions at the TBP-TATA element interface can specifically affect the response to acidic activator proteins in vivo.

This article has been cited by other articles:

• 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]  
• Larschan, E., Winston, F. (2005). The Saccharomyces cerevisiae Srb8-Srb11 Complex Functions with the SAGA Complex during Gal4-Activated Transcription. Mol. Cell. Biol. 25: 114-123 [Abstract] [Full Text]  
• Eriksson, P., Biswas, D., Yu, Y., Stewart, J. M., Stillman, D. J. (2004). TATA-Binding Protein Mutants That Are Lethal in the Absence of the Nhp6 High-Mobility-Group Protein. Mol. Cell. Biol. 24: 6419-6429 [Abstract] [Full Text]  
• Kou, H., Irvin, J. D., Huisinga, K. L., Mitra, M., Pugh, B. F. (2003). Structural and Functional Analysis of Mutations along the Crystallographic Dimer Interface of the Yeast TATA Binding Protein. Mol. Cell. Biol. 23: 3186-3201 [Abstract] [Full Text]  
• Ohdate, H., Lim, C. R., Kokubo, T., Matsubara, K., Kimata, Y., Kohno, K. (2003). Impairment of the DNA Binding Activity of the TATA-binding Protein Renders the Transcriptional Function of Rvb2p/Tih2p, the Yeast RuvB-like Protein, Essential for Cell Growth. J. Biol. Chem. 278: 14647-14656 [Abstract] [Full Text]  
• Spencer, J. V., Arndt, K. M. (2002). A TATA Binding Protein Mutant with Increased Affinity for DNA Directs Transcription from a Reversed TATA Sequence In Vivo. Mol. Cell. Biol. 22: 8744-8755 [Abstract] [Full Text]  
• Fischbeck, J. A., Kraemer, S. M., Stargell, L. A. (2002). SPN1, a Conserved Gene Identified by Suppression of a Postrecruitment-Defective Yeast TATA-Binding Protein Mutant. Genetics 162: 1605-1616 [Abstract] [Full Text]  
• Lee, M., Struhl, K. (2001). Multiple Functions of the Nonconserved N-Terminal Domain of Yeast TATA-Binding Protein. Genetics 158: 87-93 [Abstract] [Full Text]  
• Stargell, L. A., Moqtaderi, Z., Dorris, D. R., Ogg, R. C., Struhl, K. (2000). TFIIA Has Activator-dependent and Core Promoter Functions in Vivo. J. Biol. Chem. 275: 12374-12380 [Abstract] [Full Text]  
• Geisberg, J. V., Struhl, K. (2000). TATA-Binding Protein Mutants That Increase Transcription from Enhancerless and Repressed Promoters In Vivo. Mol. Cell. Biol. 20: 1478-1488 [Abstract] [Full Text]  
• Liu, Q., Gabriel, S. E., Roinick, K. L., Ward, R. D., Arndt, K. M. (1999). Analysis of TFIIA Function In Vivo: Evidence for a Role in TATA-Binding Protein Recruitment and Gene-Specific Activation. Mol. Cell. Biol. 19: 8673-8685 [Abstract] [Full Text]  
• Chou, S., Chatterjee, S., Lee, M., Struhl, K. (1999). Transcriptional Activation in Yeast Cells Lacking Transcription Factor IIA. Genetics 153: 1573-1581 [Abstract] [Full Text]  
• Shirra, M. K., Arndt, K. M. (1999). Evidence for the Involvement of the Glc7-Reg1 Phosphatase and the Snf1-Snf4 Kinase in the Regulation of INO1 Transcription in Saccharomyces cerevisiae. Genetics 152: 73-87 [Abstract] [Full Text]  
• Miyao, T., Woychik, N. A. (1998). RNA polymerase subunit RPB5 plays a role in transcriptional activation. Proc. Natl. Acad. Sci. USA 95: 15281-15286 [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]  
• Drysdale, C. M., Jackson, B. M., McVeigh, R., Klebanow, E. R., Bai, Y., Kokubo, T., Swanson, M., Nakatani, Y., Weil, P. A., Hinnebusch, A. G. (1998). The Gcn4p Activation Domain Interacts Specifically In Vitro with RNA Polymerase II Holoenzyme, TFIID, and the Adap-Gcn5p Coactivator Complex. Mol. Cell. Biol. 18: 1711-1724 [Abstract] [Full Text]  
• Kokubo, T., Swanson, M. J., Nishikawa, J.-i., Hinnebusch, A. G., Nakatani, Y. (1998). The Yeast TAF145 Inhibitory Domain and TFIIA Competitively Bind to TATA-Binding Protein. Mol. Cell. Biol. 18: 1003-1012 [Abstract] [Full Text]  
• Lehman, A. M., Ellwood, K. B., Middleton, B. E., Carey, M. (1998). Compensatory Energetic Relationships between Upstream Activators and the RNA Polymerase II General Transcription Machinery. J. Biol. Chem. 273: 932-939 [Abstract] [Full Text]  
• Mason Jr., P. B., Lis, J. T. (1997). Cooperative and Competitive Protein Interactions at the Hsp70 Promoter. J. Biol. Chem. 272: 33227-33233 [Abstract] [Full Text]  
• Hoffmann, A., Oelgeschlager, T., Roeder, R. G. (1997). Considerations of transcriptional control mechanisms: Do TFIID-core promoter complexes recapitulate nucleosome-like functions?. Proc. Natl. Acad. Sci. USA 94: 8928-8935 [Abstract] [Full Text]  
• Gonzalez-Couto, E., Klages, N., Strubin, M. (1997). Synergistic and promoter-selective activation of transcription by recruitment of transcription factors TFIID and TFIIB. Proc. Natl. Acad. Sci. USA 94: 8036-8041 [Abstract] [Full Text]  
• Kim, T. K., Roeder, R. G. (1997). Critical Role of the Second Stirrup Region of the TATA-binding Protein for Transcriptional Activation Both in Yeast and Human. J. Biol. Chem. 272: 7540-7545 [Abstract] [Full Text]  
• Nishikawa, J.-i., Kokubo, T., Horikoshi, M., Roeder, R. G., Nakatani, Y. (1997). Drosophila TAFII230 and the transcriptional activator VP16 bind competitively to the TATA box-binding domain of the TATA box-binding protein. Proc. Natl. Acad. Sci. USA 94: 85-90 [Abstract] [Full Text]  
• Bryant, G O, Martel, L S, Burley, S K, Berk, A J (1996). Radical mutations reveal TATA-box binding protein surfaces required for activated transcription in vivo.. Genes Dev. 10: 2491-2504 [Abstract]  
• Apone, L M, Virbasius, C M, Reese, J C, Green, M R (1996). Yeast TAF(II)90 is required for cell-cycle progression through G2/M but not for general transcription activation.. Genes Dev. 10: 2368-2380 [Abstract]  
• Ozer, J., Bolden, A. H., Lieberman, P. M. (1996). Transcription Factor IIA Mutations Show Activator-specific Defects and Reveal a IIA Function Distinct from Stimulation of TBP-DNA Binding. J. Biol. Chem. 271: 11182-11190 [Abstract] [Full Text]  
• Kobayashi, A., Miyake, T., Ohyama, Y., Kawaichi, M., Kokubo, T. (2001). Mutations in the TATA-binding Protein, Affecting Transcriptional Activation, Show Synthetic Lethality with the TAF145 Gene Lacking the TAF N-terminal Domain in Saccharomyces cerevisiae. J. Biol. Chem. 276: 395-405 [Abstract] [Full Text]