PUBLICATION
The human homologue of the yeast polyubiquitination factor Ufd2p is cleaved by caspase 6 and granzyme B during apoptosis
- Authors
- Mahoney, J.A., Odin, J.A., White, S.M., Shaffer, D., Koff, A., Casciola-Rosen, L., and Rosen, A.
- ID
- ZDB-PUB-020214-2
- Date
- 2002
- Source
- The Biochemical journal 361(3): 587-595 (Journal)
- Registered Authors
- Keywords
- none
- MeSH Terms
-
- Amino Acid Motifs
- Amino Acid Sequence
- Apoptosis*
- Baculoviridae/metabolism
- Binding Sites
- Blotting, Northern
- Caspase 6
- Caspases/metabolism*
- Cloning, Molecular
- Enzyme Inhibitors/pharmacology
- Fungal Proteins/chemistry*
- Fungal Proteins/metabolism
- Granzymes
- HeLa Cells
- Humans
- Kinetics
- Molecular Sequence Data
- Mutagenesis, Site-Directed
- Precipitin Tests
- Protein Binding
- Protein Structure, Tertiary
- Recombinant Proteins/metabolism
- Saccharomyces cerevisiae Proteins*
- Sequence Homology, Amino Acid
- Serine Endopeptidases/metabolism*
- Staurosporine/pharmacology
- Tissue Distribution
- Ubiquitin/metabolism
- Ubiquitin-Conjugating Enzymes
- Ultraviolet Rays
- PubMed
- 11802788 Full text @ Biochem. J.
Citation
Mahoney, J.A., Odin, J.A., White, S.M., Shaffer, D., Koff, A., Casciola-Rosen, L., and Rosen, A. (2002) The human homologue of the yeast polyubiquitination factor Ufd2p is cleaved by caspase 6 and granzyme B during apoptosis. The Biochemical journal. 361(3):587-595.
Abstract
In the present study, we demonstrate that a human homologue of Ufd2p (a yeast protein that catalyses the formation of long polyubiquitin chains, and is implicated in responses to environmental stress), UFD2 (ubiquitin fusion degradation protein-2), is cleaved during apoptosis induced by multiple stimuli, including UVB irradiation, Fas ligation, staurosporine treatment and cytotoxic lymphocyte granule-induced death. Caspase 6 and granzyme B efficiently cleave UFD2 [k(cat)/K(m)=(4-5)x10(4) M(-1).s(-1)] at Asp(123), whereas caspases 3 and 7 cleave UFD2 approx. 10-fold less efficiently immediately upstream at Asp(109). Thus UFD2 is added to the growing list of proteins with closely spaced caspase and granzyme B cleavage sites, suggesting the presence of a previously unrecognized, conserved motif. Both cleavage sites are contained and conserved within a novel 300-amino-acid N-terminal domain present in apparent UFD2 orthologues in mice and zebrafish, but absent in all UFD2 family members in lower eukaryotes. Full-length recombinant UFD2 exhibited ubiquitin-protein ligase ('E3')-like ubiquitination activity in vitro, but this activity was abolished in recombinant UFD2 truncated at the granzyme B/caspase 6 cleavage site. Cleavage of UFD2 by caspases or granzyme B within this putative regulatory N-terminal domain might have important functional consequences within the apoptotic cascade.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping