The Yeast TAF145 Inhibitory Domain and TFIIA Competitively Bind to TATA-Binding Protein

ABSTRACT The Drosophila 230-kDa TFIID subunit (dTAF230) interacts with the DNA binding domain of TATA box-binding protein (TBP) which exists in the same complex. Here, we characterize the inhibitory domain in the yeast TAF145 (yTAF145), which is homologous to dTAF230. Mutation studies show that the N-terminal inhibitory region (residues 10 to 71) can be divided into two subdomains, I (residues 10 to 37) and II (residues 46 to 71). Mutations in either subdomain significantly impair function. Acidic residues in subdomain II are important for the interaction with TBP. In addition, yTAF145 interaction is impaired by mutating the basic residues on the convex surface of TBP, which are crucial for interaction with TFIIA. Consistently, TFIIA and yTAF145 bind competitively to TBP. A deletion of the inhibitory domain of yTAF145 leads to a temperature-sensitive growth phenotype. Importantly, this phenotype is suppressed by overexpression of the TFIIA subunits, indicating that the yTAF145 inhibitory domain is involved in TFIIA function.

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The Small Nuclear RNA-activating Protein 190 Myb DNA Binding Domain Stimulates TATA Box-binding Protein-TATA Box Recognition

Journal of Biological Chemistry ◽

10.1074/jbc.m204247200 ◽

2003 ◽

Vol 278 (20) ◽

pp. 18649-18657 ◽

Cited By ~ 18

Author(s):

Craig S. Hinkley ◽

Heather A. Hirsch ◽

Liping Gu ◽

Brandon LaMere ◽

R. William Henry

Keyword(s):

Dna Binding ◽

Binding Protein ◽

Binding Domain ◽

Tata Box ◽

Dna Binding Domain ◽

Small Nuclear Rna ◽

Nuclear Rna ◽

Tata Box Binding Protein

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Functional dissection of a human Dr1-DRAP1 repressor complex.

Molecular and Cellular Biology ◽

10.1128/mcb.17.1.36 ◽

1997 ◽

Vol 17 (1) ◽

pp. 36-45 ◽

Cited By ~ 29

Author(s):

K Yeung ◽

S Kim ◽

D Reinberg

Keyword(s):

Amino Acid ◽

Dna Binding ◽

Binding Protein ◽

Transcriptional Repressor ◽

Tata Box ◽

Dna Binding Domain ◽

Specific Recognition ◽

Repressor Complex ◽

Tata Box Binding Protein ◽

Repression Domain

The heterotetrameric Dr1-DRAP1 transcriptional repressor complex was functionally dissected. Dr1 was found to contain two domains required for repression of transcription. The tethering domain interacts with the TATA box binding protein and directs the repressor complex to the promoter. This tethering domain can be replaced by a domain conferring sequence-specific recognition to the repressor complex. In the absence of the tethering domain, Dr1 interacts with its corepressor DRAP1, but this interaction is not functional. The enhancement of Dr1-mediated repression of transcription by DRAP1 requires the tethering domain. The second domain of Dr1 is the repression domain, which is glutamine-alanine rich. A 65-amino-acid polypeptide containing the repression domain fused to the Ga14 DNA binding domain repressed transcription when directed to TATA-containing and TATA-less promoters. This repression domain was also found to functionally and directly interact with the TATA box binding protein.

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Evidence that TAF-TATA Box-Binding Protein Interactions Are Required for Activated Transcription in Mammalian Cells

Molecular and Cellular Biology ◽

10.1128/mcb.22.8.2788-2798.2002 ◽

2002 ◽

Vol 22 (8) ◽

pp. 2788-2798 ◽

Cited By ~ 14

Author(s):

Lisa S. Martel ◽

Helen J. Brown ◽

Arnold J. Berk

Keyword(s):

Protein Interactions ◽

Mammalian Cells ◽

Convex Surface ◽

Binding Protein ◽

Tata Box ◽

Binding Surface ◽

Substitution Mutations ◽

Residue Substitution ◽

Tata Box Binding Protein

ABSTRACT Surfaces of human TATA box-binding protein (hsTBP) required for activated transcription in vivo were defined by constructing a library of surface residue substitution mutations and assaying them for their ability to support activated transcription in transient-transfection assays. In earlier work, three regions were identified where mutations inhibited activated transcription without interfering with TATA box DNA binding. One region is on the upstream surface of the N-terminal TBP repeat with respect to the direction of transcription and corresponds to the TBP surface that interacts with TFIIA. A second region on the stirrup of the C-terminal TBP repeat corresponds to the TFIIB-binding surface. Here we report that the third region where mutations inhibit activated transcription in mammalian cells, the convex surface of the N-terminal repeat, corresponds to a surface on TBP that interacts with hsTAF1, the major scaffold subunit of TFIID. Since mutations at the center of the hsTAF1-interacting region inhibit the ability of the protein to support activated transcription in vivo, these results are consistent with the conclusion that an interaction between hsTBP and TAFIIs is required for activated transcription in mammalian cells.

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