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Published on: March 14, 2019
Dcn1 functions as a scaffold-type E3 ligase for cullin neddylation
Thimo Kurz1, Yang-Chieh Chou, Andrew R Willems
1Institute of Biochemistry, ETH Zürich, Schafmattstrasse 18, CH-8093 Zürich, Switzerland.
Molecular Cell
|January 22, 2008
Summary
Dcn1 protein directly binds cullins and Nedd8, acting as a scaffold-like E3 ligase. This research reveals the yeast Dcn1 structure, crucial for regulating ubiquitin ligase activity via neddylation.
Area of Science:
- Molecular Biology
- Structural Biology
- Biochemistry
Background:
- Cullin-based E3 ubiquitin ligases control protein degradation and are activated by Nedd8 modification.
- Dcn1 is a key regulator of cullin neddylation and subsequent E3 ligase activity.
Purpose of the Study:
- To elucidate the structural basis of Dcn1 function in cullin neddylation.
- To characterize the interaction of Dcn1 with cullins and the Nedd8 E2 enzyme, Ubc12.
Main Methods:
- X-ray crystallography to determine the 1.9 A structure of yeast Dcn1.
- Biochemical assays to analyze Dcn1 binding and neddylation activity.
- Site-directed mutagenesis to identify key interaction surfaces.
Main Results:
- The crystal structure reveals Dcn1 comprises an N-terminal UBA domain and a novel C-terminal PONY domain.
- A conserved surface on Dcn1 mediates direct binding to cullins and is essential for neddylation.
- Dcn1 binds the Nedd8 E2 enzyme Ubc12 at a site overlapping the E1-binding region.
- Dcn1 is sufficient for cullin neddylation in a purified system, independent of cysteine residues.
Conclusions:
- Dcn1 functions as a scaffold-like E3 ligase essential for cullin neddylation.
- The structural and biochemical data provide mechanistic insights into E3 ligase regulation.
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