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P53 independent TAF6delta target genes.

ABSTRACT: To elucidate the impact of TAF6d on cell death and gene expression, here we have employed modified antisense oligonucleotides to enforce expression of endogenous TAF6d. The induction of endogenous TAF6d triggered apoptosis in tumor cell lines, including cells devoid of p53. Microarray experiments revealed that TAF6d activates gene expression independently of cellular p53 status. Our data define TAF6d as a pivotal node in a signaling pathway that acts immediately downstream of p53 to control gene expression programmes and apoptosis. Keywords: Treatment with splice site switching antisense oligonucleotides, induction of apoptosis Biological triplicates, 4 conditions: isogenic p53+/+ and -/- cells; control oligonucleotide, TAF6 AS2 oligonucleotide

ORGANISM(S): Homo sapiens

SUBMITTER: Arndt Benecke

PROVIDER: E-GEOD-10795 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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TAF6delta controls apoptosis and gene expression in the absence of p53.

Wilhelm Emmanuelle E Pellay François-Xavier FX Benecke Arndt A Bell Brendan B

PloS one 20080716 7

<h4>Background</h4>Life and death decisions of metazoan cells hinge on the balance between the expression of pro- versus anti-apoptotic gene products. The general RNA polymerase II transcription factor, TFIID, plays a central role in the regulation of gene expression through its core promoter recognition and co-activator functions. The core TFIID subunit TAF6 acts in vitro as an essential co-activator of transcription for the p53 tumor suppressor protein. We previously identified a splice varian ...[more]

PMID: 18628956

Publication: 1/2

Similar Datasets

TAF6 orchestrates transcription programs and apoptosis in the absence of p53

Project description:The decision of metazoan cells to live or undergo programmed cell death hinges on the balance between the levels of pro- versus anti-apoptotic gene products. The general RNA polymerase II (Pol II) transcription factor, TFIID, plays a central role in the regulation of gene expression through its core promoter recognition and co-activator functions. The core TFIID subunit TAF6 is a co-activator for the pro-apoptotic p53 tumour suppressor protein. Our previous studies identified a specialized isoform of TAF6, termed TAF6 that can specifically be induced in apoptosis. To elucidate the impact of TAF6 on gene expression and cell death, we employed modified antisense oligonucleotides to enforce expression of endogenous TAF6. The induction of endogenous TAF6 triggered apoptosis in several cancer cell lines. Importantly, TAF6 also induces apoptosis in tumor cell lines devoid of p53, placing TAF6 function downstream of p53. Microarray experiments uncovered a TAF6-induced transcriptome landscape displaying enhanced expression of genes of the Notch, oxidative stress, integrin, p53 and apoptosis pathways. Our data show that the TAF6 pathway is a pivotal signalling nexus that controls pro-apoptotic gene expression programs. Keywords: Treatment with splice site switching antisense oligonucleotides, induction of apoptosis Biological triplicates, 3 conditions: control oligonucleotide, Taf6 oligonucleotide, specificity control Bcl-x oligonucleotide

2010-11-27 | E-GEOD-8752 | biostudies-arrayexpress

Modulation of gene expression in U251 glioblastoma cells by binding of mutant p53 to intronic and intergenic sequences

Project description:Missense point mutations in the TP53 gene are frequent genetic alterations in human tumor tissue and cell lines derived thereof. Mutant p53 (mutp53) proteins have lost sequence-specific DNA binding, but have retained the ability to interact in a structure-selective manner with non-B DNA and to act as regulators of transcription. To identify functional binding sites of mutp53, we established a small library of genomic sequences bound by p53R273H in U251 human glioblastoma cells using chromatin immunoprecipitation (ChIP). Mutp53 binding to isolated DNA fragments confirmed the specificity of the ChIP. The mutp53 bound DNA sequences are rich in repetitive DNA elements, which are dispersed over non-coding DNA regions. Stable down-regulation of mutp53 expression strongly suggested that mutp53 binding to genomic DNA is functional. We identified the PPARGC1A and FRMD5 genes as p53R273H targets regulated by binding to intronic and intra-genic sequences. We propose a model that attributes the oncogenic functions of mutp53 to its ability to interact with intronic and intergenic non-B DNA sequences and modulate gene transcription via re-organization of chromatin. For the study of the consequences of mutant p53 (R273H) knockdown on gene expression, total RNA from parental U251 glioblastoma cells and UsiA12 clone was prepared from two independent cell culture experiments (biological replicates) and processed for microarray-based profiling of gene expression. UsiA12 clone was derived from the U251 cells transfected with the pSuper-p53 and pCI-neo vectors.

2010-06-24 | E-GEOD-13991 | biostudies-arrayexpress

Determining the impact of alternative splicing events on transcriptome dynamics

Project description:Background The complete sequencing of the human genome and its subsequent analysis revealed a predominant role for alternative splicing in the generation of proteome diversity. Splice switching oligonucleotides (SSOs) are a powerful and specific tool to experimentally control alternative splicing of endogenous messenger RNAs in living cells. SSOs also have therapeutic potential to treat diseases that are caused by aberrant splicing. The assignment of biological roles to alternative splicing events of currently unknown function promises to provide a largely untapped source of potential new therapeutic targets. Here we have developed a protocol that combines high sensitivity microarrays with the transfection of SSOs to monitor global changes in gene expression downstream of alternate, endogenous splice events. Findings When applied to a well-characterized splicing event in the Bcl-x gene, the application of high sensitivity microarrays revealed a link between the induction of the Bcl-xS isoform and the repression of genes involved in protein synthesis. Conclusions The strategy introduced herein provides a useful approach to define the biological impact of any given alternative splicing event on global gene expression patterns. Furthermore, our data provide the first link between Bcl-xS expression and the repression of ribosomal protein gene expression. Biological triplicates, 2 conditions: control oligonucleotide, Bcl-x oligonucleotide

2008-10-02 | E-GEOD-9851 | biostudies-arrayexpress

Gene expression profiling of human anaplastic thyroid carcinoma cells by microarray

Project description:In this study, we profiled the change of gene expression by p53 knock-down on anaplastic thyroid carcinoma (ATC) cell line, KAT-18 cells using Applied Biosystem's Microarrays. When compared with the untreated control KAT-18 cells, 2,926 genes were found to have altered expression levels of 2-fold or more in cells treated with antisense p53 oligonucleotide. Microarray analysis revealed that p53 knock-down modified the expression of numerous apoptosis-related genes. Among those genes, 33 apoptosis pathway activators and 20 apoptosis pathway repressors were identified. We analyzed human thyroid carcinoma cell (KAT18) on Applied Biosystem Human 1700 Chip Arrays. There were control and test sample, respectively, KAT18 untreated and treated antisense p53.

2010-06-25 | E-GEOD-12912 | biostudies-arrayexpress

Gene expression profile in pelvic organ prolapse

Project description:Introduction and Hypothesis: Identify processes contributing to pelvic organ prolapse (POP) by transcriptional profiling of pelvic connective tissue in conjunction with light microscopy. Methods: We performed a frequency matched case-control study of women undergoing hysterectomy. Total RNA, extracted from uterosacral and round ligament samples used to generate labeled cRNA, was hybridized to microarrays and analyzed for the expression of 32,878 genes. Significance Analysis of Microarrays, (Stanford University, CA), identified differentially expressed genes used for ontoanalysis, and quantitative PCR (qPCR) confirmed results. Light microscopy confirmed tissue type and assessed inflammatory infiltration. Results: The analysis of thirty-four arrays revealed 249 differentially expressed genes with fold changes larger than 1.5 fold and false discovery rates M-bM-^IM-$5.2%. Immunity and Defense was the most significant biological process differentially expressed in POP. Selected qPCR confirmed 4 genes. Light microscopy showed no inflammatory infiltrates. Conclusions: Genes enriched for Immunity and Defense contribute to POP independent of inflammatory infiltrates. Keywords: whole tissue (endopelvic fascia) type comparison This was a group matched case control study of 8 women with pelvic organ prolapse versus 9 non-prolapse controls, both undergoing hysterectomy for benign conditions. Two separate pelvic support tissues were collected from each patient. The uterosacral ligament and round ligament tissue was removed at the time of hysterectomy, RNA was extracted and ABI whole genome chips used to identify differences in expression profiles of individual samples. Various ethnic groups, age groups and menopausal status were included.

2008-09-19 | E-GEOD-12852 | biostudies-arrayexpress

EBER2 RNA-induced transcriptome changes identify cellular processes likely targeted during Epstein Barr Virus infection.

Project description:Little is known about the physiological role of the EBER1 and 2 nuclear RNAs during Epstein Barr viral infection. The cellular transcription response to EBER2 expression using the wild-type and an internal deletion mutant was determined. Significant changes in gene expression patterns were observed. A functional meta-analysis of the regulated genes points to inhibition of stress and immune responses, as well as activation of cellular growth and cytoskeletal reorganization as potential targets for EBER2 RNA. Different functions can be assigned to different parts of the RNA. These results provide new avenues to the understanding of EBER2 and EBV biology. Keywords: Overexpression of non-translated RNA Biological relicates, 3 conditions: empty expression vector (pUC-18), pUC-18-EBER2, pUC-18-EBER2-L2

2008-10-02 | E-GEOD-11093 | biostudies-arrayexpress

Effect of fetal HDL on gene expression in human placental endothelial cells

Project description:The aim of this study was to identify HDL and apoE-regulated genes in human placental endothelial cells (HPEC), which are exposed to fetal HDL. HPECs extracted from 5 human placentas were cultivated and treated for 16 h with 15ug/ml fetal HDL, 15ug/ml reconstituted HDL (rHDL), or endothelial basal medium (EBM) as vehicle control. Gene expression analysis from these 3 conditions (5 biological replicates) using 15 Applied Biosystems Human Whole Genome Survey V2.0 Microarrays was perfomed and significantly differentially expressed genes between two different groups (HDL vs control or rHDL vs control) were identified.

2011-09-01 | E-MEXP-3299 | biostudies-arrayexpress

The Applied Biosystems Tissue Gene Expression Database - Human Body Map. A whole genome survey of 32 Human Tissues

Project description:The Applied Biosystems Tissue Gene Expression Database, known as the Human Body Map, is a collection of gene expression profiles from 31 normal tissues and a Universal Human Reference RNA (Stratagene). The Human BodyMap was created using the Applied Biosystems Expression Array System. There were three replicates per tissue for a total of 96 microarrays. Keywords: Human tissue whole genome survey 31 human tissues from Clontech and 1 from Stratagene (UHR) were used to create the Applied Biosystems Human BodyMap (a.k.a.Tissue Gene Expression Database ). Digoxigenin-UTP-labeled cRNA was generated and linearly amplified from 1 µg of total RNA with three replicates per tissue, using the Applied Biosystems Chemiluminescent RT-IVT Labeling Kit v 2.0 and kit protocol. Microarray hybridization, chemiluminescence detection, image acquisition, and analysis were performed on the on the Applied Biosystems 1700 Chemiluminescent Microarray Analyzer using the Applied Biosystems Chemiluminescence Detection Kit and kit protocol. This dataset is part of the TransQST collection.

2007-06-04 | E-GEOD-7905 | biostudies-arrayexpress

Transcription profiling of human cell types after 24 hrs serum starvation

Project description:Comparison of cell types after 24 hrs serum starvation.

2006-05-01 | E-MEXP-519 | biostudies-arrayexpress

Differential Gene Expression Between Brown Norway and Sprague Dawley Rats in Response to Renal Ischemia Reperfusion

Project description:Renal ischemia reperfusion injury (I/R) has great clinical relevance and many treatments and hospitalizations are associated with this pathological process. Animal models are widely used in the development of kidney I/R studies and numerous papers have been published in this field in recent years suggesting that the nature of the animal species used in these models can lead to a different response to ischemic insult. The Brown Norway (BN) rat strain is known to show endogenous resistance to the ischemic process and several studies have sought to identify the cell mechanism present in this resistance. Despite of all the previously reported works, little is know about the genetic response of BN in front of ischemic insult. By means of a microarray comparative genetic study between BN and the ischemia susceptible rat strain Sprague Dawley (SD), the present work determine the genetic response to the I/R process in both rat strains and the genes that are differentially expressed during the I/R process and link them with some of the main cytoprotective processes responsible for the endogenous resistance to renal ischemic injury in BN rats. The following experimental groups for BN and SD rats (n=3 per group) were studied: Group 1: Control (C): Animals subjected to identical maneuvers as the I/R group except that the renal pedicles were not clamped. Group 2: Ischemia/Reperfusion (I/R): Animals were subjected to 45 min of bilateral ischemia and 24 h of reperfusion. Gene expression profiles were obtained from control and I/R groups in the two rat strains in three biological replicates, resulting in 12 hybridizations from 2 groups × 2 strains × 3biological replicates. To evaluate the technical reliability of microarrays, replicate microarrays were performed on several RNA samples. In total, 12 hybridizations were performed for this study.

2007-12-27 | E-GEOD-9943 | biostudies-arrayexpress

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