title: The E3 ubiquitin ligase Ro52 negatively regulates IFN-beta production post-pathogen recognition by polyubiquitin-mediated degradation of IRF3.
abstract: Induction of type I IFNs is a fundamental cellular response to both viral and bacterial infection. The role of the transcription factor IRF3 is well established in driving this process. However, equally as important are cellular mechanisms for turning off type I IFN production to limit this response. In this respect, IRF3 has previously been shown to be targeted for ubiquitin-mediated degradation postviral detection to turn off the IFN-beta response. In this study, we provide evidence that the E3 ligase Ro52 (TRIM21) targets IRF3 for degradation post-pathogen recognition receptor activation. We demonstrate that Ro52 interacts with IRF3 via its C-terminal SPRY domain, resulting in the polyubiquitination and proteasomal degradation of the transcription factor. Ro52-mediated IRF3 degradation significantly inhibits IFN-beta promoter activity, an effect that is reversed in the presence of the proteasomal inhibitor MG132. Specific targeting of Ro52 using short hairpin RNA rescues IRF3 degradation following polyI:C-stimulation of HEK293T cells, with a subsequent increase in IFN-beta production. Additionally, shRNA targeting of murine Ro52 enhances the production of the IRF3-dependent chemokine RANTES following Sendai virus infection of murine fibroblasts. Collectively, this demonstrates a novel role for Ro52 in turning off and thus limiting IRF3-dependent type I IFN production by targeting the transcription factor for polyubiquitination and subsequent proteasomal degradation.
authors.affiliation: Molecular and Cellular Therapeutics, Research Institute, Royal College of Surgeons in Ireland, Dublin, Ireland.
authors.forename: Rowan
authors.initials: R
authors.lastname: Higgs
authors.forename: Joan
authors.initials: J
authors.lastname: Ní Gabhann
authors.forename: Nadia
authors.initials: N
authors.lastname: Ben Larbi
authors.forename: Eamon P
authors.initials: EP
authors.lastname: Breen
authors.forename: Katherine A
authors.initials: KA
authors.lastname: Fitzgerald
authors.forename: Caroline A
authors.initials: CA
authors.lastname: Jefferies
mesh_terms: D002460:Cell Line; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases / Q000378:metabolism; D006801:Humans; D050838:Interferon Regulatory Factor-3 / Q000235:genetics / Q000378:metabolism*; D016899:Interferon-beta / Q000096:biosynthesis* / Q000235:genetics; D025821:Polyubiquitin / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D011971:Receptors, Immunologic; D012261:Ribonucleoproteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D015398:Signal Transduction; D051196:Toll-Like Receptor 3 / Q000378:metabolism; D051197:Toll-Like Receptor 4 / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D000099207:SS-A Antigen
chemical_list: D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases; D050838:Interferon Regulatory Factor-3; D016899:Interferon-beta; D025821:Polyubiquitin; D011971:Receptors, Immunologic; D012261:Ribonucleoproteins; D051196:Toll-Like Receptor 3; D051197:Toll-Like Receptor 4; D044767:Ubiquitin-Protein Ligases; D000099207:SS-A Antigen; C494232:IRF3 protein, human; C495344:TLR3 protein, human; C495345:TLR4 protein, human; C452820:RIGI protein, human