RNA-seq identified GLI1 as the target gene for GABP
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ABSTRACT:
PROVIDER: PRJNA1024439 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR26297647_1.fastq.gz | Fastqsanger.gz | |||
| SRR26297647_2.fastq.gz | Fastqsanger.gz | |||
| SRR26297648_1.fastq.gz | Fastqsanger.gz | |||
| SRR26297648_2.fastq.gz | Fastqsanger.gz | |||
| SRR26297649_1.fastq.gz | Fastqsanger.gz |
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Project description:We performed RNA-seq based transcriptomic analysis to evaluate differential protein expression in the kidneys of GABP overexpressing mice in db/m and GABP knockdown mice in db/db. The cluster analysis heat map shows the genetic changes . Compared with db/m mice, 734 differentially expressed genes, 525 upregulated genes and 209 upregulated genes . Compared with db/db mice, 82 genes were up-regulated and 82 genes were down-regulated in the kidneys of mice with GABP knockdown in db/db . Further, the classical pathway map in IPA database found that cell growth, proliferation, organ development and other pathways were activated after GABP overexpression. Cell growth, proliferation, organ development and other pathways were inhibited . It was further confirmed that GABP was associated with proliferation. Among differentially expressed genes, there were 9 differentially expressed genes up-regulated by GABP overexpression and down-regulated by GABP knockdown . Through the analysis of Ingenuity Pathway Analysis database, GABP and GLI1 were most closely related .
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Project description:Panc-1GLI1ER and Panc-1ER were established from human pancreatic cancer cell line Panc-1. Panc-1GLI1ER cells express a chimeric transgene composed of an enhanced green fluorescent protein (EGFP)-tagged version of the amino-terminal half of GLI1, fused to the AF2 domain of the mouse estrogen receptor 1(Esr1) cDNA. On the other hand, Panc-1ER harbor only EGFP-tagged AF2 domain. To identify GLI1 direct target genes, total RNAs were purified from the cells before and 3 hours after the beta-estradiol (E2) treatment. Gene expression profiles were analyzed by AGILENT human 4x44 cDNA microarray. Keywords: Expression profiling by genome tiling array GLI1 direct target genes were identified in Panc-1 cells. Gene expression profiles of Panc-1GLI1ER and Panc-1ER cells were analyzed by AGILENT human 4x44 cDNA microarray before and 3 hours after treatment of beta-estradiol (E2) at doses of 10nM.
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Project description:GLI1 is a transcription factor correlated to decreased survival in several cancers. We have identified SMARCA2 as a co-regulator that enhances GLI1-mediated transcriptional activity and functions through the C-terminal transcriptional activation domain of GLI1. Central domains including the ATPase motif of SMARCA2 physically interact with GLI1. Evaluation of DNA density indicates GLI1, like SMARCA2, can increase the DNA accessibility with a preference for sites distal to gene transcription start sites and outside the promoter regions (i.e. enhancers). The putative enhancers where accessibility is decreased by the knock down of GLI1 and SMARCA2 are located cis to genes, such as HHIP, that are regulated by GLI1 and implicated in cancer functions. At the putative enhancer for HHIP, the localization of SMARCA2 is at least partially dependent on GLI1’s presence. Understanding this transcriptional regulation by GLI1 and SMARCA2 through altering chromatin accessibility at enhances can provide additional therapeutic targets for cancers dependent on GLI1.
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Project description:We knocked out different exons corresponding to different sections of GABP, a transcription factor known to bind to the telomerase reverse transcriptase (TERT) promoter with the G228 mutation. We investigated the effects of the knockouts in the regulation of TERT expression and other subunits of GABP. GABP genomic binding sites were determined through chromatin immunoprecipitation sequencing (ChIP-seq), and gene expression was determined through total RNA sequencing (RNA-seq). Combining data pertaining to GABP binding sites and gene expression provided insight into the molecular mechanisms of maintaining the length of telomeres in cancers.
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Project description:GLI1 is a transcription factor correlated to decreased survival in several cancers. We have identified SMARCA2 as a co-regulator that enhances GLI1-mediated transcriptional activity and functions through the C-terminal transcriptional activation domain of GLI1. Central domains including the ATPase motif of SMARCA2 physically interact with GLI1. Evaluation of DNA density indicates GLI1, like SMARCA2, can increase the DNA accessibility with a preference for sites distal to gene transcription start sites and outside the promoter regions (i.e. enhancers). The putative enhancers where accessibility is decreased by the knock down of GLI1 and SMARCA2 are located cis to genes, such as HHIP, that are regulated by GLI1 and implicated in cancer functions. At the putative enhancer for HHIP, the localization of SMARCA2 is at least partially dependent on GLI1’s presence. Understanding this transcriptional regulation by GLI1 and SMARCA2 through altering chromatin accessibility at enhances can provide additional therapeutic targets for cancers dependent on GLI1.
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Project description:In each cell type the expression of genes is regulated by the action of a large number of transcription factors, but so far we have only a rudimentary knowledge of the location of the gene regulatory elements where they bind. This can now be addressed with genome-wide ChIP experiments. In a previous ChIP-chip study of USF1 and USF2 we found evidence also of binding of GABP, FOXA2 and HNF4a within the enriched regions. Here we have applied ChIP-seq for these transcription factors and identified 3064 peaks of enrichment for GABP, 7266 for FOXA2 and 18783 for HNF4a. HNF4a and FOXA2 binding was found in at least half of the regions previously identified as bound by USF2 but not USF1, showing that they frequently bind the same regulatory elements. GABP peaks were found at transcription start sites whereas 94 % of FOXA2 and 90 % of HNF4a peaks were located at other positions. We developed a method based on the high resolution achieved by ChIP-seq to accurately define TFBS within peaks, and found the predicted sites to have an elevated conservation level compared to peak centers; however the majority of bindings were not evolutionary conserved. An unexpected interaction between HNF4a and GABP was seen at TSS, with as many as 1/3 of the HNF4a positive promoters being bound also by GABP, and co-immunoprecipitations show that these factors are in the same complex in the nucleus. ArrayExpress Release Date: 2009-06-10 Publication Title: Molecular interactions between HNF4a, FOXA2 and GABP identified at regulatory DNA elements through ChIP-sequencing Publication Author List: Ola Wallerman, Mehdi Motallebipour, Stefan Enroth, Kalicharan Patra, MadhuSudan Reddy Bysani, Jan Komorowski, Claes Wadelius Person Roles: submitter Person Last Name: Wallerman Person First Name: Ola Person Mid Initials: Person Email: ola.wallerman@genpat.uu.se Person Phone: Person Address: IGP, Rudbecklab Person Affiliation: Uppsala University
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