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PMID:15485925
| Citation |
Kim, KI, Giannakopoulos, NV, Virgin, HW and Zhang, DE (2004) Interferon-inducible ubiquitin E2, Ubc8, is a conjugating enzyme for protein ISGylation. Mol. Cell. Biol. 24:9592-600 |
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| Abstract |
Protein ISGylation is unique among ubiquitin-like conjugation systems in that the expression and conjugation processes are induced by specific stimuli, mainly via the alpha/beta interferon signaling pathway. It has been suggested that protein ISGylation plays a special role in the immune response, because of its interferon-signal dependency and its appearance only in higher eukaryotic organisms. Here, we report the identification of an ISG15-conjugating enzyme, Ubc8. Like other components of the protein ISGylation system (ISG15, UBE1L, and UBP43), Ubc8 is an interferon-inducible protein. Ubc8 clearly mediates protein ISGylation in transfection assays. The reduction of Ubc8 expression by small interfering RNA causes a decrease in protein ISGylation in HeLa cells upon interferon treatment. Neither UbcH7/UbcM4, the closest homologue of Ubc8 among known ubiquitin E2s, nor the small ubiquitin-like modifier E2 Ubc9 supports protein ISGylation. These findings strongly suggest that Ubc8 is a major ISG15-conjugating enzyme responsible for protein ISGylation upon interferon stimulation. Furthermore, we established an assay system to detect ISGylated target proteins by cotransfection of ISG15, UBE1L, and Ubc8 together with a target protein to be analyzed. This method provides an easy and effective way to identify new targets for the ISGylation system and will facilitate related studies. |
| Links |
PubMed PMC522249 Online version:10.1128/MCB.24.21.9592-9600.2004 |
| Keywords |
Animals; Base Sequence; Cells, Cultured; Cytokines/chemistry; Cytokines/genetics; Cytokines/metabolism; DNA-Binding Proteins/metabolism; Humans; Interferons/pharmacology; Mice; Molecular Sequence Data; Promoter Regions, Genetic/genetics; Protein Binding; Protein Processing, Post-Translational; RNA Interference; STAT1 Transcription Factor; Signal Transduction/drug effects; Trans-Activators/metabolism; Two-Hybrid System Techniques; Ubiquitin-Activating Enzymes/deficiency; Ubiquitin-Activating Enzymes/genetics; Ubiquitin-Activating Enzymes/metabolism; Ubiquitin-Conjugating Enzymes/chemistry; Ubiquitin-Conjugating Enzymes/genetics; Ubiquitin-Conjugating Enzymes/metabolism; Ubiquitins/analogs & derivatives; Ubiquitins/chemistry; Ubiquitins/genetics; Ubiquitins/metabolism |
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Significance
Annotations
| Gene product | Qualifier | GO ID | GO term name | Evidence Code | with/from | Aspect | Notes | Status |
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References
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