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PMID: 15485925 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Interferon-inducible ubiquitin E2, Ubc8, is a conjugating enzyme for protein ISGylation.

Molecular and cellular biology ·Vol. 24 ·No. 21 ·2004-11-00 ·Pages 9592-600

Kim KI, Giannakopoulos NV, Virgin HW, Zhang DE

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.

MeSH Terms
Animals Base Sequence Cells, Cultured Cytokines/chemistry,genetics,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,genetics,metabolism Ubiquitin-Conjugating Enzymes/chemistry,genetics,metabolism Ubiquitins/analogs & derivatives,chemistry,genetics,metabolism
Chemicals
Cytokines DNA-Binding Proteins STAT1 Transcription Factor STAT1 protein, human Stat1 protein, mouse Trans-Activators Ubiquitins ISG15 protein, human Interferons Ubiquitin-Conjugating Enzymes ubiquitin-conjugating enzyme UBC8 Ubiquitin-Activating Enzymes
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Kim Keun Il
The Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA 92037, USA.
Giannakopoulos Nadia V
Virgin Herbert W
Zhang Dong-Er
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.24.21.9592-9600.2004
Published
2004-11-00
Pages
9592-600
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC522249
Subset
IM
Grants
NIGMS NIH HHS · R01 GM066955 · United States
NCI NIH HHS · CA079849 · United States
NCI NIH HHS · R01 CA079849 · United States
NIAID NIH HHS · U54 AI057160 · United States
NIGMS NIH HHS · GM066955 · United States
External Links
PubMed source DOI: 10.1128/MCB.24.21.9592-9600.2004 PMC Full Text View journal details
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