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

2007 ◽  
Vol 28 (1) ◽  
pp. 83-92 ◽  
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.

scholarly journals Taenia solium TAF6 and TAF9 binds to a putative Downstream Promoter Element present in TsTBP1 gene core promoter

2017 ◽  
Author(s):  
Oscar Rodríguez-Lima ◽  
Ponciano García-Gutierrez ◽  
Lucía Jimenez ◽  
Angel Zarain-Herzberg ◽  
Roberto Lazarini ◽  
...  
Keyword(s):  
Molecular Model ◽  
Binding Protein ◽  
Core Promoter ◽  
Taenia Solium ◽  
Tata Binding Protein ◽  
Tata Box ◽  
Promoter Element ◽  
Core Promoters ◽  
Downstream Promoter Element ◽  
The Core

AbstractWe have cloned and characterized the gene encoding to Taenia solium TATA binding protein 1 (TsTBP1). It spans 1481 bp and its coding region is interrupted by four introns that possess the consensus donor/acceptor sequences. It produces a protein of 238 amino acids residues, which presented all the classical motives of the TBP1. On the core promoter region we identified putative binding sites for NF1, AP-1, YY1, TAF1/TAF2 and TAF6/TAF9, and the TSS that corresponds to an A+1. Southern and Northern blot analysis showed that TsTBP1 is encoded by a single gene, which produce a messenger of about 1.1 kbp with a higher differential expression in adult than the larval stage. Moreover, two putative TATA boxes were identified at -97 and -69 bp (relative to the TSS), likewise a Downstream Promoter Element (DPE) was located at +27 to +31 bp. EMSA experiments did not show any component of cysticerci nuclear extracts bound to the putative TATA-box elements; in contrast TAF6 and TAF9 bind to the DPE, showing that TsTBP1 is a TATA-less gen. On the other hand, TAF6 and TAF9 were localized on the nucleus from cells of T. crassiceps cysticerci bladder walls. By using the amino acid sequences of T. solium TAF6 (TsTAF6) and TAF9 (TsTAF9), we constructed a molecular model showing interaction between DPE-TsTAF6 and TsTAF6-TsTAF9. Finally, an in silico analysis of the core promoters of Taeniidae family genes showed that TATA-box and DPE are homologous to the mammalian elements, but not so the Inr. The low identity between TAF9 from cestodes and mammals open the possibility to use it as target to interrupt or modulate the transcription of TATA-box less genes in cestodes as a novel therapeutic strategy.Author summaryNeurocysticercosis still being a health problem in developing countries. Mexico reports a prevalence of 2.5% in the total patients from National Institute of Neurology and Neurosurgery. Several efforts have been made in different fields to study Taenia solium, and although the Genome Project has been published and released the genomic sequence of this parasite; the transcriptional mechanisms this organism remains unexplored. We isolated and characterized Taenia solium TATA-Binding Protein 1 (TsTBP1) gene and TBP-associated factor 6 and 9 cDNAs (TsTAF6 and TsTAF9, respectively). Our principal findings are: 1.- Identification of cis and trans elements in the core promoter of TsTBP1 gene. 2.- The TsTBP1 gene is a TATA-less promoter. 3.- Cloning of cDNAs to TsTAF6 and TsTAF9, 4.- Identification of a DPE in the core promoter of TsTBP1 gene, which interacts with TAF6 and TAF9, 5.- Construction of a molecular model that shows the interaction between DPE, TAF6, and TAF9; and 6.- A proposal of consensus sequences for TATA-box, Inr and DPE presented on Taeniidae family core promoters.

scholarly journals Adeno-Associated Virus Major Rep78 Protein Disrupts Binding of TATA-Binding Protein to the p97 Promoter of Human Papillomavirus Type 16

2000 ◽  
Vol 74 (5) ◽  
pp. 2459-2465 ◽  
Author(s):  
Pei-Fen Su ◽  
Shu-Yuan Chiang ◽  
Cheng-Wen Wu ◽  
Felicia Y.-H. Wu
Keyword(s):  
Human Papillomavirus ◽  
Binding Protein ◽  
Core Promoter ◽  
Tata Binding Protein ◽  
Tata Box ◽  
Dependent Manner ◽  
Hpv 16 ◽  
Adeno Associated Virus ◽  
Human Papillomavirus Type 16 ◽  
E6 And E7

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.

Affinity, stability and polarity of binding of the TATA binding protein governed by flexure at the TATA box 1 1Edited by P. E. Wright

1998 ◽  
Vol 282 (4) ◽  
pp. 731-739 ◽  
Author(s):  
Anne Grove ◽  
Aldo Galeone ◽  
Elaine Yu ◽  
Luciano Mayol ◽  
E.Peter Geiduschek
Keyword(s):  
Binding Protein ◽  
Tata Binding Protein ◽  
Tata Box

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

2014 ◽  
Vol 35 (5) ◽  
pp. 601-608 ◽  
Author(s):  
Irina Drachkova ◽  
Ludmila Savinkova ◽  
Tatyana Arshinova ◽  
Mikhail Ponomarenko ◽  
Sergey Peltek ◽  
...  
Keyword(s):  
Binding Protein ◽  
Tata Binding Protein ◽  
Hereditary Diseases ◽  
Tata Box

scholarly journals Functional Analysis of the Three TATA Binding Protein Homologs in Methanosarcina acetivorans

2010 ◽  
Vol 192 (6) ◽  
pp. 1511-1517 ◽  
Author(s):  
Matthew J. Reichlen ◽  
Katsuhiko S. Murakami ◽  
James G. Ferry
Keyword(s):  
Binding Protein ◽  
Growth Yield ◽  
Tata Binding Protein ◽  
Lag Phase ◽  
Methanosarcina Acetivorans ◽  
Wild Type ◽  
Molecular Approaches ◽  
Lag Times ◽  
Microarray Analyses ◽  
Expression Strain

ABSTRACT The roles of three TATA binding protein (TBP) homologs (TBP1, TBP2, and TBP3) in the archaeon Methanosarcina acetivorans were investigated by using genetic and molecular approaches. Although tbp2 and tbp3 deletion mutants were readily obtained, a tbp1 mutant was not obtained, and the growth of a conditional tbp1 expression strain was tetracycline dependent, indicating that TBP1 is essential. Transcripts of tbp1 were 20-fold more abundant than transcripts of tbp2 and 100- to 200-fold more abundant than transcripts of tbp3, suggesting that TBP1 is the primary TBP utilized during growth. Accordingly, tbp1 is strictly conserved in the genomes of Methanosarcina species. Δtbp3 and Δtbp2 strains exhibited an extended lag phase compared with the wild type, although the lag phase for the Δtbp2 strain was less pronounced when this strain was transitioning from growth on methylotrophic substrates to growth on acetate. Acetate-adapted Δtbp3 cells exhibited growth rates, final growth yields, and lag times that were significantly reduced compared with those of the wild type when the organisms were cultured with growth-limiting concentrations of acetate, and the acetate-adapted Δtbp2 strain exhibited a final growth yield that was reduced compared with that of the wild type when the organisms were cultured with growth-limiting acetate concentrations. DNA microarray analyses identified 92 and 77 genes with altered transcription in the Δtbp2 and Δtbp3 strains, respectively, which is consistent with a role for TBP2 and TBP3 in optimizing gene expression. Together, the results suggest that TBP2 and TBP3 are required for efficient growth under conditions similar to the conditions in the native environment of M. acetivorans.

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

1995 ◽  
Vol 9 (23) ◽  
pp. 2974-2985 ◽  
Author(s):  
S K Whitehall ◽  
G A Kassavetis ◽  
E P Geiduschek
Keyword(s):  
Binding Protein ◽  
Tata Binding Protein ◽  
Tata Box ◽  
U6 Rna

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

1998 ◽  
Vol 18 (7) ◽  
pp. 3771-3781 ◽  
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.

Sign in / Sign up

Export Citation Format

Share Document