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| Status |
Public on May 14, 2014 |
| Title |
Hypoxia regulates alternative splicing of HIF and non-HIF target genes |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Alternative RNA splicing analysis in Hep3B cell cultured under 21% (N1,3,5) or 1.2% (H2,4,6) oxygen
Hypoxia is a common characteristic of many solid tumors. The hypoxic microenvironment stabilizes hypoxia-inducible transcription factor 1α (HIF1α) and 2α (HIF2α) to activate gene transcription, which promotes tumor cell survival. 95% of human genes are alternatively spliced, producing RNA isoforms that code functionally distinct proteins. Thus, effective hypoxia response requires increased HIF target gene transcription as well as proper RNA splicing of these HIF target genes. However, it is unclear if and how hypoxia regulates RNA splicing of HIF target genes. This study determined the effects of hypoxia on alternative splicing (AS) of HIF and non-HIF target genes in Hep3B cells and characterized the role of HIF in regulating AS of HIF induced genes. The results indicated that hypoxia generally promotes exon inclusion for hypoxia-induced, but reduces exon inclusion for hypoxia reduced genes. Mechanistically, HIF activity, but not hypoxia per se is found to be necessary and sufficient to increase exon inclusion of several HIF target genes including pyruvate dehydrogenase kinase 1 (PDK1). PDK1 splicing reporters confirmed that transcriptional activation by HIF is sufficient to increase exon inclusion of PDK1 splicing reporter. In contrast, transcriptional activation of the PDK1 minigene by other transcription factor in the absence of endogenous HIF target gene activation fails to alter PDK1 RNA splicing, demonstrating a novel role of HIF target gene(s) in regulating RNA splicing of HIF target genes. Implications:This study demonstrates a novel function of HIF in regulating RNA splicing of HIF target genes.
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| Overall design |
We analyzed total RNA from Hep3B cells cultured under 21% (N1,3,5) or 1.2% (H2,4,6) oxygen using the Affymetrix Human Exon 1.0 ST platform. Array data was processed by Altanalyze software version 2.0.7. Techinical replicates were performed for Nx and Hx treated Hep3B cells
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| Contributor(s) |
Sena J, Hu C, Wang L, Heasley L |
| Citation(s) |
24850901 |
| Submission date |
May 13, 2014 |
| Last update date |
Aug 13, 2014 |
| Contact name |
Cheng-Jun Hu |
| E-mail(s) |
cheng-jun.hu@ucdenver.edu
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| Phone |
3037244574
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| Organization name |
University of Colorado Denver
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| Department |
Craniofacial Biology
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| Lab |
Cheng-Jun Hu
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| Street address |
E. 17th Ave
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| City |
Aurora |
| State/province |
CO |
| ZIP/Postal code |
80045 |
| Country |
USA |
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| Platforms (1) |
| GPL10520 |
[HuEx-1_0-st] Affymetrix Human Exon 1.0 ST Array [HuEx-1_0-st-v2,mainR3,A20071112,EP.cdf] |
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| Samples (6)
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| GSM1385550 |
Hep3B Cells 21% Oxygen, biological rep1 |
| GSM1385551 |
Hep3B Cells 21% Oxygen, biological rep2 |
| GSM1385552 |
Hep3B Cells 21% Oxygen, biological rep3 |
| GSM1385553 |
Hep3B Cells 1.5% Oxygen, biological rep1 |
| GSM1385554 |
Hep3B Cells 1.5% Oxygen, biological rep2 |
| GSM1385555 |
Hep3B Cells 1.5% Oxygen, biological rep3 |
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| Relations |
| BioProject |
PRJNA246960 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE57613_RAW.tar |
132.4 Mb |
(http)(custom) |
TAR (of CEL) |
| Processed data included within Sample table |
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