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Using In Vitro Fluorescence Resonance Energy Transfer to Study the Dynamics Of Protein Complexes at a Millisecond Time Scale
Published on: March 14, 2019
DEN1 deneddylates non-cullin proteins in vivo
Yaru Chan1, Jeongsook Yoon, June-Tai Wu
1Institute of Molecular Biology, Academia Sinica, 128 Sec No. 2 Academia Road, Taipei 115, Taiwan.
Journal of Cell Science
|September 11, 2008
Summary
The cysteine protease DEN1/NEDP1 removes ubiquitin-like protein Nedd8/Rub1 from cellular proteins. DEN1 is crucial for animal viability by maintaining neddylation and deneddylation balance.
Area of Science:
- Biochemistry
- Molecular Biology
- Cellular Biology
Background:
- Ubiquitin-like protein Nedd8/Rub1 activates cullin ubiquitin ligases through covalent modification.
- The full range of Nedd8-modified proteins and the regulation of protein neddylation remain unclear.
Purpose of the Study:
- To characterize the Drosophila DEN1 protein and its role in protein neddylation.
- To investigate the deneddylation activity of DEN1 in vitro and in vivo.
Main Methods:
- Characterization of Drosophila DEN1 protein and DEN1 null mutants.
- In vitro deneddylation assays using purified DEN1 protein.
- In vivo analysis of protein neddylation status in DEN1 mutants.
Main Results:
- DEN1 processes the Nedd8 precursor and deneddylates numerous cellular proteins, not just cullins.
- Purified DEN1 efficiently deneddylates Cul1 and Cul3, but their neddylated levels are not elevated in DEN1 null mutants.
- DEN1 null mutants exhibit widespread protein hyper-neddylation, which is reversed by purified DEN1.
- DEN1's deneddylation activity is distinct from the CSN complex.
Conclusions:
- DEN1 plays a significant role in deneddylation beyond cullin proteins.
- A balance between neddylation and deneddylation, regulated by DEN1, is essential for animal survival.
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