X-ray crystal structure of IRF-3 and its functional implications

Takahasi, Kiyohiro; Suzuki, Nobuo; Horiuchi, Masataka; Mori, Mitsuaki; Suhara, Wakako; Okabe, Yasutaka; Fukuhara, Yukiko; Terasawa, Hiroaki; Akira, Shizuo; Fujita, Takashi; Inagaki, Fuyuhiko

doi:10.1038/nsb1001

Cited by 151 publications

(158 citation statements)

References 34 publications

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“…Recently, we determined the crystal structure of IRF-3-(175-427), which retains the minimum structure needed for virusinduced phosphorylation and dimerization (25). Although the crystallized IRF-3-(175-427) was not phosphorylated, it formed a dimer.…”

Section: Resultsmentioning

confidence: 99%

Identification of Ser-386 of Interferon Regulatory Factor 3 as Critical Target for Inducible Phosphorylation That Determines Activation

Mori¹,

Yoneyama²,

Ito

et al. 2004

Journal of Biological Chemistry

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196

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Interferon regulatory factor (IRF)-3 is a critical transcription factor regulating innate immune responses against viral and bacterial infections. Signals activated by various pathogens are integrated by IRF-3 kinase, resulting in the specific phosphorylation of IRF-3 in the cytoplasm. This phosphorylation induces dimerization and association with the coactivators CREB-binding protein/p300, and the resultant complex activates the target genes in the nucleus. However, the phosphorylation sites that determine the active/inactive status of IRF-3 have been a source of controversy. In this study, we generated an antibody that specifically detects the phosphorylation of Ser-386 and used it as a probe. We found: 1) viral infection specifically induces phosphorylation of the Ser-386; 2) recently identified IRF-3 kinases (IKK-i/⑀ and TBK-1) phosphorylate Ser-386 and induce its dimerization; 3) phosphorylation of Ser-386 is exclusively observed with the dimer; 4) mutation at Ser-386 abolishes the dimerization potential; 5) a constitutively active 5D mutant designed to mimic phosphorylation of Ser/Thr residues other than Ser-385 and -386 is secondarily phosphorylated at Ser-386, presumably by an irrelevant kinase. These results strongly suggest that Ser-386 is the target of the IRF-3 kinase and critical determinant for the activation of IRF-3.

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Section: Resultsmentioning

confidence: 99%

Identification of Ser-386 of Interferon Regulatory Factor 3 as Critical Target for Inducible Phosphorylation That Determines Activation

Mori¹,

Yoneyama²,

Ito

et al. 2004

Journal of Biological Chemistry

Self Cite

196

185

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“…It is the dimerized form of IRF3 that activates type I IFN responses (Takahasi et al, 2003). A knockdown of TRIM21 increased levels of IRF3 dimers in IFN-γ-treated cells stimulated with double-stranded DNA (Herring testis [HT]-DNA) transfected into the cells (Fig.…”

Section: Trim21 Is a Regulator Receptor For Autophagic Degradation Ofmentioning

confidence: 99%

“…Detection of IRF3 dimerization was performed by native PAGE as previously described (Takahasi et al, 2003). Quantitative RT-PCR was performed as previously described using the following primer sets: ULK1, 5′-AGATGTTCCAGCACCGTGAG and AATGCACAGCTTGCACTTGG-3′; BECN1, 5′-GGAGAACCT-CAGCCGAAGAC-3′ and 5′-ACGTTGAGCTGAGTGTCCAG-3′; ACTIN, 5′-GGGCATGGGTCAGAAGGATT-3′ and 5′-TCGATGG-GGTACTTCAGGGT-3′; TRIM1, 5′-AAGAATGTGACGAGTTG-GTAGAG-3′ and ATGAGGACTGTTGACCGTTC-3′; TRIM5, 5′-CATGCCTCACTGCAAACCAC-3′ and 5′-GGTAACTGATCCG-GCACACA-3′; TRIM8, 5′-ATCCTGATGGACAGGACCCA-3′ and 5′-CTCCTTCTTGGCCACTTCGT-3′; TRIM16, 5′-GTAAGCCCAC-GAACACAAATG-3′ and 5′-TCCAGCCCTGAAACTTCTATTC-3′; TRIM20, 5′-CTGAGTCAGGAGCACCAAGG-3′ and 5′-GCT-GCTCCTCCCCTGATTTT-3′; TRIM21, 5′-CAGTCTCGGAAACAC-CGTGA-3′ and 5′-AATGCCACCTGGAGCTTCTC-3′; TRIM22, 5′-CTCGACCTGCTTATCCGTATTT-3′ and 5′-CTCAGCACAAGG-GCTACTATG-3′; TRIM28, 5′-CCATACTGTGCGCTCTACTG-3′ and 5′-GGTTCATGCTTGTGTACGTTG-3′; TRIM56, 5′-TTCTTCGT-CAATGGGCTGCT-3′ and 5′-AAGTCATCGGCACAGTCCAG-3′; and TRIM65, 5′-GATCTACCTGAACTTGCCTCTG-3′ and 5′-GAG-GAGGGAGGAATCTGTCT-3′.…”

Section: Irf3 Dimerization Assay and Quantitative Rt-pcrmentioning

confidence: 99%

TRIM-mediated precision autophagy targets cytoplasmic regulators of innate immunity

Kimura¹,

Jain²,

Choi³

et al. 2015

J Exp Med

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“…19,20 Since IRF3-CL and IRF3 share a common amino terminus, we investigated whether IRF3-CL could form a homo-or a heterodimer with IRF3 constitutively. It was clear that Flag-tagged IRF3-CL was co-immunoprecipitated with Myc-tagged IRF3-CL but not with Myc-tagged IRF3 (Figure 6a), thus indicating the association between IRF3-CL molecules, but not between IRF3-CL and IRF3.…”

Section: Irf3-cl An Isoform Of Irf3 Antagonizes Activity Of Irf3 Chmentioning

confidence: 99%

Interferon regulatory factor 3-CL, an isoform of IRF3, antagonizes activity of IRF3

Chen

2010

Cell Mol Immunol

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Interferon regulatory factor 3 (IRF3), one member of the IRF family, plays a central role in induction of type I interferon (IFN) and regulation of apoptosis. Controlled activity of IRF3 is essential for its functions. During reverse transcription (RT)-PCR to clone the full-length open reading frame (ORF) of IRF3, we cloned a full-length ORF encoding an isoform of IRF3, termed as IRF3-CL, and has a unique carboxyl-terminus of 125 amino acids. IRF3-CL is ubiquitously expressed in distinct cell lines. Overexpression of IRF3-CL inhibits Sendai virus (SeV)-triggered induction of IFN-b and SeV-induced and inhibitor of NF-kB kinase-e (IKKe)-mediated nuclear translocation of IRF3. When IKKe is overexpressed, IRF3-CL is associated with IRF3. These results suggest that IRF3-CL, the alternative splicing isoform of IRF-3, may function as a negative regulator of IRF3. Keywords: interferon regulatory factor 3; negative regulation; splicing variant INTRODUCTIONThe interferon regulatory factor (IRF) family of transcriptional factors plays versatile roles in many biological processes, including innate and adaptive immune responses, cell growth control, apoptosis and hematopoietic development. 1,2 To date, nine members of the mammalian IRF family have been identified and characterized, each containing a highly conserved DNA-binding domain of ,120 amino acids (aa) at the amino terminus, and a variable carboxyl-terminal IRF association domain (IAD). 2 IRF3, one member of the IRF family, possesses a transactivation domain (aa 134-394) and two autoinhibition domains. 3 After viral infection, the latent IRF3, which resides primarily in the cytoplasm, is phosphorylated by the inhibitor of NF-kB kinase-e (IKKe) and TANK-binding kinase 1 (TBK1) and forms a homo-or heterodimer with other transcriptional factors which then translocates into the nucleus. [4][5][6] The activated IRF3 associates with the transcriptional cofactors cyclic AMP-responsive elementbinding protein and P300 to bind to the specific DNA targets, thus leading to transcriptional initiation of target genes with other cofactors simultaneously recruited. 7 Accumulating evidence demonstrates that IRF3 plays an essential role in the virus-or bacterium-mediated induction of interferon (IFN)-b and a subset of IFN-stimulated genes through the Toll-like receptors or the cytosolic receptors pathway. 8-10 Furthermore, IRF3 has been shown to function as a vital signaling regulator in the innate immune response, development of immune cells and apoptosis. 11 As rapid and controlled cellular responses are pivotal to host defense, the activity of IRF3 should be regulated at multiple levels.

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X-ray crystal structure of IRF-3 and its functional implications

Cited by 151 publications

References 34 publications

Identification of Ser-386 of Interferon Regulatory Factor 3 as Critical Target for Inducible Phosphorylation That Determines Activation

Identification of Ser-386 of Interferon Regulatory Factor 3 as Critical Target for Inducible Phosphorylation That Determines Activation

TRIM-mediated precision autophagy targets cytoplasmic regulators of innate immunity

Interferon regulatory factor 3-CL, an isoform of IRF3, antagonizes activity of IRF3

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