Abstract
The interferon regulatory factor (IRF)-3 transcription factor plays a central role in the capacity of the host to mount an efficient innate antiviral immune defense, mainly through the regulation of type I Interferon genes. A tight regulation of IRF-3 is crucial for an adapted intensity and duration of the response. Redox-dependent processes are now well known to regulate signaling cascades. Recent reports have revealed that signaling molecules upstream of IRF-3, including the mitochondrial antiviral-signalling protein (MAVS) and the TNF receptor associated factors (TRAFs) adaptors, are sensitive to redox regulation. In the present study, we assessed whether redox regulation of thiol residues contained in IRF-3, which are priviledged redox sensors, play a role in its regulation following Sendai virus infection, using a combination of mutation of Cysteine (Cys) residues into Alanine and thiols alkylation using N-ethyl maleimide. Alkylation of IRF-3 on Cys289 appears to destabilize IRF-3 dimer in vitro. However, a detailed analysis of IRF-3 phosphorylation, dimerization, nuclear accumulation, and induction of target gene promoter in vivo led us to conclude that IRF-3 specific, individual Cys residues redox status does not play an essential role in its activation in vivo.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Active Transport, Cell Nucleus / genetics
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Active Transport, Cell Nucleus / immunology
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Adaptor Proteins, Signal Transducing / genetics
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Adaptor Proteins, Signal Transducing / immunology
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Adaptor Proteins, Signal Transducing / metabolism
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Alkylation
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Amino Acid Substitution
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Animals
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Cell Line, Transformed
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Cell Nucleus / genetics
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Cell Nucleus / immunology
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Cell Nucleus / metabolism*
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Ethylmaleimide / chemistry
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Interferon Regulatory Factor-3 / genetics
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Interferon Regulatory Factor-3 / immunology
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Interferon Regulatory Factor-3 / metabolism*
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Interferon Type I / biosynthesis
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Interferon Type I / genetics
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Interferon Type I / immunology
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Mice
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Mice, Knockout
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Mutation, Missense
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Promoter Regions, Genetic / genetics
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Promoter Regions, Genetic / immunology
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Protein Multimerization*
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Respirovirus Infections / genetics
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Respirovirus Infections / immunology
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Respirovirus Infections / metabolism*
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Sendai virus / immunology
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Sendai virus / metabolism*
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Tumor Necrosis Factor Receptor-Associated Peptides and Proteins / genetics
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Tumor Necrosis Factor Receptor-Associated Peptides and Proteins / immunology
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Tumor Necrosis Factor Receptor-Associated Peptides and Proteins / metabolism
Substances
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Adaptor Proteins, Signal Transducing
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IPS-1 protein, mouse
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Interferon Regulatory Factor-3
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Interferon Type I
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Irf3 protein, mouse
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Tumor Necrosis Factor Receptor-Associated Peptides and Proteins
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Ethylmaleimide