 |
 |
GEO help: Mouse over screen elements for information. |
|
| Status |
Public on Sep 13, 2024 |
| Title |
EAPP is essential for tri-snRNP biogenesis and controls cell fate and tumor growth via the alternative splicing of MDM4 |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
Precursor messenger RNA (pre-mRNA) splicing is catalyzed by the spliceosome, a highly dynamic machinery with sequentially assembled small nuclear ribonucleoproteins (snRNPs) and splicing factors. Aberrant spliceosome composition and function result in the dysregulation of pre-mRNA alternative splicing, which may cause cellular stresses and diseases such as cancer. Here, we identify a splicing factor, E2F-associated phosphoprotein (EAPP), that directly binds to the U4/U6. U5 tri-snRNP and PRPF19 complex. Notably, the C-terminal “bucket” domain in EAPP composed of several beta-sheets is required for its interaction with the tri-snRNP. EAPP is prominently located at Cajal bodies within the nucleus where it colocalizes with the spliceosome. Loss of EAPP impedes the assembly and homeostasis of the tri-snRNP complex in Cajal bodies and increases the frequency of splicing abnormalities, mostly exon skipping. Moreover, EAPP depletion promotes MDM4 exon 6 skipping, which suppresses cell growth and tumor progression via p53 overactivation. Our study demonstrates a previously uncharacterized function of EAPP in spliceosome regulation and reveals that EAPP-mediated alternative splicing of MDM4 is a critical determinant of cell fate and tumor growth.
|
| |
|
| Overall design |
To further investigate how EAPP affects the downstream regulatory mechanisms of tumor cell growth, we utilized CRISPR/Cas9 technology to knock out EAPP in HeLa cells. Infect cells with lentiCRISPR-Cas9 lentivirus. After 48 hours, add 2 μg/ml puromycin to select for transduced cells, and select and expand single-cell clones.
We then performed gene expression profiling analysis using data obtained from RNA-seq. Comparative gene expression and alternative splicing profiling analysis of RNA-seq data for EAPP knockout and WT HeLa cells.
|
| |
|
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
| |
| Submission date |
Sep 09, 2024 |
| Last update date |
Sep 13, 2024 |
| Contact name |
Haiping Zhao |
| Organization name |
Fudan University
|
| Street address |
131 DongAn Road
|
| City |
Shanghai |
| ZIP/Postal code |
200032 |
| Country |
China |
| |
|
| Platforms (1) |
| GPL24676 |
Illumina NovaSeq 6000 (Homo sapiens) |
|
| Samples (6)
|
|
| Relations |
| BioProject |
PRJNA1158679 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE276681_EAPP_alternative_splicing_events.xlsx |
57.8 Kb |
(ftp)(http) |
XLSX |
| GSE276681_EAPP_differentially_expressed_genes.xlsx |
266.4 Kb |
(ftp)(http) |
XLSX |
| GSE276681_EAPP_gene_counts.xlsx |
6.2 Mb |
(ftp)(http) |
XLSX |
| GSE276681_EAPP_gene_fpkm.xlsx |
7.8 Mb |
(ftp)(http) |
XLSX |
SRA Run Selector |
| Raw data are available in SRA |
|
|
|
|
 |
Less...
More...