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| Status |
Public on Jan 02, 2019 |
| Title |
Fbxo32 mediated gene expression program underlies EMT and metastasis |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
The epithelial-mesenchymal transition (EMT) is a process by which cells lose their cell contacts and gain migratory and invasive properties. EMT is essential for numerous developmental processes including neural tube formation, in wound healing, organ fibrosis and cancer metastasis. Despite progress, the repertoire of factors involved in global transcriptional reprogramming underlying EMT remains unknown. Here we show that FBXO32, a member of the F-box protein family, is essential for phenotypic changes hallmark of EMT. Such dependency results from FBXO32-driven transcriptional reprogramming of critical EMT genes and involved changes in their chromatin state. Furthermore, we found that CTBP1, an established regulator of EMT, requires FBXO32 ubiquitination at Lysine 63 for its nuclear retention and gene regulatory function. FBXO32 is also highly amplified in a large panel of metastatic cancers and its knockdown severely impaired metastatic properties of cancer cells in vitro and in vivo. In addition, FBXO32 is also induced during neurogenesis and is essential for neuronal migration in vivo. Together, these findings uncover FBXO32-dependent gene regulatory circuitry that underlies EMT during development and disease.
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| Overall design |
For all siRNA mediated knockdown experiments, cells were seeded at same starting density and transfected with ON-TARGET plus SMART pool siRNAs (i.e. a mixture of 4 siRNA provided as a single reagent) (Dharmacon) every second day. For siRNA transfections, Lipofectamine RNAiMax (Invitrogen, 13778-150) was used according to the manufacturer's instructions. For experiments during TGF-β-induced EMT, TGF-β-induction performed at the same time when siRNA was added to avoid indirect effect due to loss of function of protein.
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| Contributor(s) |
Sahu SK, Pataskar A, Tiwari V |
| Citation missing |
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| |
| Submission date |
Jan 29, 2016 |
| Last update date |
May 15, 2019 |
| Contact name |
Abhijeet Pataskar |
| Organization name |
Institute of Molecular Biology
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| Street address |
Ackermannweg 4
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| City |
Mainz |
| ZIP/Postal code |
55128 |
| Country |
Germany |
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| Platforms (1) |
| GPL11154 |
Illumina HiSeq 2000 (Homo sapiens) |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA310235 |
| SRA |
SRP069086 |
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