 |
 |
GEO help: Mouse over screen elements for information. |
|
| Status |
Public on Aug 15, 2009 |
| Title |
The epithelial splicing factors ESRP1 and ESRP2 positively and negatively regulate alternative splicing events |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
|
| Summary |
Epithelial Splicing Regulatory Proteins 1 and 2 (ESRP1 and ESRP2) are recently discovered epithelial-specific RNA-binding proteins that promote splicing of the epithelial variant of the FGFR2, ENAH, CD44, and CTNND1 transcripts. To catalogue a larger set of splicing events under the regulation of the ESRPs, we profiled splicing changes induced by RNA interference-mediated knockdown of ESRP1 and ESRP2 expression in a human epithelial cell line using the splicing-sensitive Affymetrix Exon ST1.0 Arrays. Analysis of the microarray data using the previously described MADS tool resulted in the identification of over a hundred candidate ESRP-regulated splicing events. We were able to independently validate 37 of these targets by RT-PCR. The ESRP-regulated events encompass all known types of alternative splicing events. Importantly, a number of these regulated splicing events occur in gene transcripts that encode proteins with well-described roles in the regulation of actin cytoskeleton organization, cell-cell adhesion, cell polarity, and cell migration. In sum, this work reveals a novel list of transcripts differentially spliced in epithelial and mesenchymal cells, implying that coordinated alternative splicing plays a critical role in determination of cell type identity.
Keywords: control / knockdown comparison
|
| |
|
| Overall design |
Short interfering knockdown of ESRP1 and ESRP2 in human PNT2 prostatic epithelium cells was performed as described before (Warzecha et al., 2009, Molecular Cell 33:591-601). The efficiency of ESRP1 and ESRP2 knockdown was monitored by quantitative RT-PCR as described before (Warzecha et al., 2009, Molecular Cell 33:591-601). In all cases the efficiency of the knockdown was close to 80%. We conducted Exon array profiling on RNAs from four siESRP1/2-treated samples and four siGFP controls.
|
| |
|
| Contributor(s) |
Warzecha CC, Shen S, Xing Y, Carstens RP |
| Citation(s) |
19829082 |
| Submission date |
Aug 02, 2009 |
| Last update date |
Feb 18, 2019 |
| Contact name |
Yi Xing |
| E-mail(s) |
yi-xing@uiowa.edu
|
| Phone |
319-384-3099
|
| Fax |
319-384-3150
|
| Organization name |
University of Iowa
|
| Department |
Internal Medicine, Biomedical Engineering
|
| Street address |
3294 CBRB, 285 Newton Rd
|
| City |
Iowa City |
| State/province |
IA |
| ZIP/Postal code |
52242 |
| Country |
USA |
| |
|
| Platforms (1) |
| GPL5175 |
[HuEx-1_0-st] Affymetrix Human Exon 1.0 ST Array [transcript (gene) version] |
|
| Samples (8)
|
|
| Relations |
| BioProject |
PRJNA118751 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE17468_RAW.tar |
178.1 Mb |
(http)(custom) |
TAR (of CEL) |
| Processed data included within Sample table |
|
|
|
|
 |
Less...
More...