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PMID: 19854139 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Key role of Ubc5 and lysine-63 polyubiquitination in viral activation of IRF3.

Molecular cell ·Vol. 36 ·No. 2 ·2009-10-23 ·Pages 315-25

Zeng W, Xu M, Liu S, Sun L, Chen ZJ

Abstract

The mitochondrial antiviral signaling protein (MAVS; also known as IPS-1, VISA, and CARDIF) is essential for innate immune response against RNA viruses. MAVS transduces signals from the cytosolic RIG-I-like receptors, which bind to viral RNAs. But how MAVS activates downstream transcription factors such as IRF3 to induce type-I interferons is not well understood. We have established a cell-free system in which mitochondria derived from virus-infected cells activate IRF3 in the cytosol. Fractionation of the cytosol led to the identification of Ubc5 as a ubiquitin-conjugating enzyme (E2) required for IRF3 activation. Using an inducible RNAi strategy, we demonstrate that catalytically active Ubc5 is required for IRF3 activation by viral infection. The activation of IRF3 also requires two ubiquitin-binding domains of NEMO. Furthermore, we show that replacement of endogenous ubiquitin with its K63R mutant abolishes viral activation of IRF3, demonstrating that K63 polyubiquitination plays a key role in IRF3 activation.

MeSH Terms
Amino Acid Sequence Biocatalysis Cell Fractionation HeLa Cells Humans I-kappa B Kinase/chemistry,metabolism Interferon Regulatory Factor-3/metabolism Lysine/metabolism Molecular Sequence Data Polyubiquitin/metabolism Protein Structure, Tertiary Sendai virus/physiology Ubiquitin-Conjugating Enzymes/chemistry,metabolism Ubiquitination Virus Activation
Chemicals
IKBKG protein, human IRF3 protein, human Interferon Regulatory Factor-3 Polyubiquitin UBE2D1 protein, human Ubiquitin-Conjugating Enzymes I-kappa B Kinase Lysine
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Zeng Wenwen
Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390-9148, USA.
Xu Ming
Liu Siqi
Sun Lijun
Chen Zhijian J
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Article Info
Journal
Molecular cell
Abbr.
Mol Cell
ISSN
1097-4164
DOI
10.1016/j.molcel.2009.09.037
Published
2009-10-23
Pages
315-25
Language
English
Region
United States
NLM ID
9802571
PMCID
PMC2779157
Subset
IM
Grants
NIGMS NIH HHS · R01-GM63692 · United States
NIAID NIH HHS · R01 AI060919 · United States
NIAID NIH HHS · F32 AI009919 · United States
Howard Hughes Medical Institute · United States
NIAID NIH HHS · R01-AI09919 · United States
NIGMS NIH HHS · R01 GM063692-08 · United States
NIGMS NIH HHS · R01 GM063692 · United States
NIAID NIH HHS · R01 AI060919-05 · United States
External Links
PubMed source DOI: 10.1016/j.molcel.2009.09.037 PMC Full Text View journal details
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