Electrochemical impedance probing of transcriptional TATA binding protein based on TATA box site-specific binding

ABSTRACT Adeno-associated virus type 2 (AAV) is known to inhibit the promoter activities of several oncogenes and viral genes, including the human papillomavirus type 16 (HPV-16) E6 and E7 transforming genes. However, the target elements of AAV on the long control region (LCR) upstream of E6 and E7 oncogenes are elusive. A chloramphenicol acetyltransferase assay was performed to study the effect of AAV on the transcription activity of the HPV-16 LCR in SiHa (HPV-positive) and C-33A (HPV-negative) cells. The results reveal that (i) AAV inhibited HPV-16 LCR activity in a dose-dependent manner, (ii) AAV-mediated inhibition did not require the HPV gene products, and (iii) the AAV replication gene product Rep78 was involved in the inhibition. Deletion mutation analyses of the HPV-16 LCR showed that regulatory elements outside the core promoter region of the LCR may not be direct targets of AAV-mediated inhibition. Further study with the electrophoretic mobility shift assay demonstrated that Rep78 interfered with the binding of TATA-binding protein (TBP) to the TATA box of the p97 core promoter more significantly than it disrupted the preformed TBP-TATA complex. These data thus suggest that Rep78 may inhibit transcription initiation of the HPV-16 LCR by disrupting the interaction between TBP and the TATA box of the p97 core promoter.

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The Mechanism by which TATA-Box Polymorphisms Associated with Human Hereditary Diseases Influence Interactions with the TATA-Binding Protein

Human Mutation ◽

10.1002/humu.22535 ◽

2014 ◽

Vol 35 (5) ◽

pp. 601-608 ◽

Cited By ~ 21

Author(s):

Irina Drachkova ◽

Ludmila Savinkova ◽

Tatyana Arshinova ◽

Mikhail Ponomarenko ◽

Sergey Peltek ◽

...

Keyword(s):

Binding Protein ◽

Tata Binding Protein ◽

Hereditary Diseases ◽

Tata Box

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Variations in Intracellular Levels of TATA Binding Protein Can Affect Specific Genes by Different Mechanisms

Molecular and Cellular Biology ◽

10.1128/mcb.00809-07 ◽

2007 ◽

Vol 28 (1) ◽

pp. 83-92 ◽

Cited By ~ 4

Author(s):

Stephanie D. Bush ◽

Patricia Richard ◽

James L. Manley

Keyword(s):

Binding Protein ◽

Core Promoter ◽

Tata Binding Protein ◽

Tata Box ◽

Wild Type ◽

The Core ◽

Mitotic Delay ◽

Reduced Activity ◽

Dt40 Cells ◽

Repressor Element

ABSTRACT We previously showed that reduced intracellular levels of the TATA binding protein (TBP), brought about by tbp heterozygosity in DT40 cells, resulted in a mitotic delay reflecting reduced expression of the mitotic regulator cdc25B but did not significantly affect overall transcription. Here we extend these findings in several ways. We first provide evidence that the decrease in cdc25B expression reflects reduced activity of the cdc25B core promoter in the heterozygous (TBP-het) cells. Strikingly, mutations in a previously described repressor element that overlaps the TATA box restored promoter activity in TBP-het cells, supporting the idea that the sensitivity of this promoter to TBP levels reflects a competition between TBP and the repressor for DNA binding. To determine whether cells might have mechanisms to compensate for fluctuations in TBP levels, we next examined expression of the two known vertebrate TBP homologues, TLP and TBP2. Significantly, mRNAs encoding both were significantly overexpressed relative to levels observed in wild-type cells. In the case of TLP, this was shown to reflect regulation of the core promoter by both TBP and TLP. Together, our results indicate that variations in TBP levels can affect the transcription of specific promoters in distinct ways, but overall transcription may be buffered by corresponding alterations in the expression of TBP homologues.

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The symmetry of the yeast U6 RNA gene's TATA box and the orientation of the TATA-binding protein in yeast TFIIIB.

Genes & Development ◽

10.1101/gad.9.23.2974 ◽

1995 ◽

Vol 9 (23) ◽

pp. 2974-2985 ◽

Cited By ~ 50

Author(s):

S K Whitehall ◽

G A Kassavetis ◽

E P Geiduschek

Keyword(s):

Binding Protein ◽

Tata Binding Protein ◽

Tata Box ◽

U6 Rna

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Even-skipped Represses Transcription by Binding TATA Binding Protein and Blocking the TFIID-TATA Box Interaction

Molecular and Cellular Biology ◽

10.1128/mcb.18.7.3771 ◽

1998 ◽

Vol 18 (7) ◽

pp. 3771-3781 ◽

Cited By ~ 38

Author(s):

Chi Li ◽

James L. Manley

Keyword(s):

Rna Polymerase Ii ◽

Binding Protein ◽

Direct Interaction ◽

In Vitro Transcription ◽

Tata Binding Protein ◽

Tata Box ◽

Homeodomain Protein ◽

Necessary And Sufficient

ABSTRACT The Drosophila homeodomain protein Even-skipped (Eve) is a transcriptional repressor, and previous studies have suggested that it functions by interfering with the basal transcription machinery. Here we describe experiments indicating that the mechanism of Eve repression involves a direct interaction with the TATA binding protein (TBP) that blocks binding of TBP-TFIID to the promoter. We first compared Eve activities in in vitro transcription systems reconstituted with either all the general transcription factors or only TBP, TFIIB, TFIIF30, and RNA polymerase II. In each case, equivalent and very efficient levels of repression were observed, indicating that no factors other than those in the minimal system are required for repression. We then show that Eve can function efficiently when its recognition sites are far from the promoter and that the same regions of Eve required for repression in vivo are necessary and sufficient for in vitro repression. This includes, in addition to an Ala-Pro-rich region, residues within the homeodomain. Using GAL4-Eve fusion proteins, we demonstrate that the homeodomain plays a role in repression in addition to DNA binding, which is to facilitate interaction with TBP. Single-round transcription experiments indicate that Eve must function prior to TBP binding to the promoter, suggesting a mechanism whereby Eve represses by competing with the TATA box for TBP binding. Consistent with this, excess TATA box-containing oligonucleotide is shown to specifically and efficiently disrupt the TBP-Eve interaction. Importantly, we show that Eve binds directly to TFIID and that this interaction can also be disrupted by the TATA oligonucleotide. We conclude that Eve represses transcription via a direct interaction with TBP that blocks TFIID binding to the promoter.

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

Molecular and Cellular Biology ◽

10.1128/mcb.18.2.1003 ◽

1998 ◽

Vol 18 (2) ◽

pp. 1003-1012 ◽

Cited By ~ 93

Author(s):

Tetsuro Kokubo ◽

Mark J. Swanson ◽

Jun-ichi Nishikawa ◽

Alan G. Hinnebusch ◽

Yoshihiro Nakatani

Keyword(s):

Convex Surface ◽

Binding Protein ◽

Tata Binding Protein ◽

Tata Box ◽

Temperature Sensitive ◽

Dna Binding Domain ◽

Inhibitory Region ◽

Inhibitory Domain ◽

Growth Phenotype ◽

Tata Box 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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