Cleavage of the C-terminus of NEDD8 by UCH-L3.
NEDD8 is a novel ubiquitin-like protein that has been shown to conjugate to nuclear proteins in a manner analogous to ubiquitination and sentrinization. To identify proteins that are involved in the NEDD8-conjugation and de-conjugation pathway, the yeast two-hybrid system was used to screen a human heart cDNA library using NEDD8 as a bait. Seven strongly positive clones were found to contain a cDNA insert encoding the ubiquitin C-terminal hydrolase, UCH-L3. In vitro GST pull-down assay demonstrated that UCH-L3 bound to both NEDD8 and ubiquitin. In contrast, UCH-L3 did not bind to sentrin-1, sentrin-2, or sentrin-3. Recombinant UCH-L3, but not UCH-L1, was able to cleave the C-terminus of NEDD8. Thus, UCH-L3 can function as a C-terminal hydrolase for both NEDD8 and ubiquitin. UCH-L3 may play a physiologically significant role in the cleavage of the C-terminus of NEDD8, which is required for NEDD8 to conjugate to target proteins.
Wada
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Division of Molecular Medicine, Department of Internal Medicine, and Research Center for Cardiovascular Diseases, Institute of Molecular Medicine for the Prevention of Human Diseases, The University of Texas- Houston Health Science Center, Houston,
Kito
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Caskey
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Yeh
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Kamitani
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D015536:Down-Regulation; D015723:Gene Library; D006801:Humans; D006868:Hydrolysis; D000075687:NEDD8 Protein; D010446:Peptide Fragments / Q000378:metabolism; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D012441:Saccharomyces cerevisiae / Q000235:genetics; D012641:Selection, Genetic; D013869:Thiolester Hydrolases / Q000235:genetics / Q000378:metabolism*; D014018:Tissue Distribution; D043222:Ubiquitin Thiolesterase; D014452:Ubiquitins / Q000235:genetics / Q000378:metabolism*
D000075687:NEDD8 Protein; C082860:NEDD8 protein, human; D010446:Peptide Fragments; D014452:Ubiquitins; D013869:Thiolester Hydrolases; D043222:Ubiquitin Thiolesterase