Synthetic mimetics assigned a major role to IFNAR2 in type I interferon signaling
Type I interferons (IFNs) are potent inhibitors of viral replication. Here, we reformatted the natural murine and human type I interferon-α/β receptors IFNAR1 and IFNAR2 into fully synthetic biological switches. The transmembrane and intracellular domains of natural IFNAR1 and IFNAR2 were conserved,...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-09-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2022.947169/full |
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| author | Nele Zoellner Noémi Coesfeld Frederik Henry De Vos Jennifer Denter Haifeng C. Xu Elena Zimmer Birgit Knebel Hadi Al-Hasani Sofie Mossner Philipp A. Lang Doreen M. Floss Jürgen Scheller |
| author_facet | Nele Zoellner Noémi Coesfeld Frederik Henry De Vos Jennifer Denter Haifeng C. Xu Elena Zimmer Birgit Knebel Hadi Al-Hasani Sofie Mossner Philipp A. Lang Doreen M. Floss Jürgen Scheller |
| author_sort | Nele Zoellner |
| collection | DOAJ |
| description | Type I interferons (IFNs) are potent inhibitors of viral replication. Here, we reformatted the natural murine and human type I interferon-α/β receptors IFNAR1 and IFNAR2 into fully synthetic biological switches. The transmembrane and intracellular domains of natural IFNAR1 and IFNAR2 were conserved, whereas the extracellular domains were exchanged by nanobodies directed against the fluorescent proteins Green fluorescent protein (GFP) and mCherry. Using this approach, multimeric single-binding GFP-mCherry ligands induced synthetic IFNAR1/IFNAR2 receptor complexes and initiated STAT1/2 mediated signal transduction via Jak1 and Tyk2. Homodimeric GFP and mCherry ligands showed that IFNAR2 but not IFNAR1 homodimers were sufficient to induce STAT1/2 signaling. Transcriptome analysis revealed that synthetic murine type I IFN signaling was highly comparable to IFNα4 signaling. Moreover, replication of vesicular stomatitis virus (VSV) in a cell culture-based viral infection model using MC57 cells was significantly inhibited after stimulation with synthetic ligands. Using intracellular deletion variants and point mutations, Y510 and Y335 in murine IFNAR2 were verified as unique phosphorylation sites for STAT1/2 activation, whereas the other tyrosine residues in IFNAR1 and IFNAR2 were not involved in STAT1/2 phosphorylation. Comparative analysis of synthetic human IFNARs supports this finding. In summary, our data showed that synthetic type I IFN signal transduction is originating from IFNAR2 rather than IFNAR1. |
| first_indexed | 2024-04-14T02:02:44Z |
| format | Article |
| id | doaj.art-a844187356564f8ba83cf36bff594e71 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-04-14T02:02:44Z |
| publishDate | 2022-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-a844187356564f8ba83cf36bff594e712022-12-22T02:18:47ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2022-09-011310.3389/fmicb.2022.947169947169Synthetic mimetics assigned a major role to IFNAR2 in type I interferon signalingNele Zoellner0Noémi Coesfeld1Frederik Henry De Vos2Jennifer Denter3Haifeng C. Xu4Elena Zimmer5Birgit Knebel6Hadi Al-Hasani7Sofie Mossner8Philipp A. Lang9Doreen M. Floss10Jürgen Scheller11Medical Faculty, Institute of Biochemistry and Molecular Biology II, Heinrich-Heine-University, Düsseldorf, GermanyMedical Faculty, Institute of Biochemistry and Molecular Biology II, Heinrich-Heine-University, Düsseldorf, GermanyMedical Faculty, Institute of Biochemistry and Molecular Biology II, Heinrich-Heine-University, Düsseldorf, GermanyMedical Faculty, Institute of Molecular Medicine II, Heinrich-Heine-University, Düsseldorf, GermanyMedical Faculty, Institute of Molecular Medicine II, Heinrich-Heine-University, Düsseldorf, GermanyMedical Faculty, Institute of Biochemistry and Molecular Biology II, Heinrich-Heine-University, Düsseldorf, GermanyMedical Faculty, Institute for Clinical Biochemistry and Pathobiochemistry, German Diabetes Center, Heinrich-Heine-University, Düsseldorf, GermanyMedical Faculty, Institute for Clinical Biochemistry and Pathobiochemistry, German Diabetes Center, Heinrich-Heine-University, Düsseldorf, GermanyMedical Faculty, Institute of Biochemistry and Molecular Biology II, Heinrich-Heine-University, Düsseldorf, GermanyMedical Faculty, Institute of Molecular Medicine II, Heinrich-Heine-University, Düsseldorf, GermanyMedical Faculty, Institute of Biochemistry and Molecular Biology II, Heinrich-Heine-University, Düsseldorf, GermanyMedical Faculty, Institute of Biochemistry and Molecular Biology II, Heinrich-Heine-University, Düsseldorf, GermanyType I interferons (IFNs) are potent inhibitors of viral replication. Here, we reformatted the natural murine and human type I interferon-α/β receptors IFNAR1 and IFNAR2 into fully synthetic biological switches. The transmembrane and intracellular domains of natural IFNAR1 and IFNAR2 were conserved, whereas the extracellular domains were exchanged by nanobodies directed against the fluorescent proteins Green fluorescent protein (GFP) and mCherry. Using this approach, multimeric single-binding GFP-mCherry ligands induced synthetic IFNAR1/IFNAR2 receptor complexes and initiated STAT1/2 mediated signal transduction via Jak1 and Tyk2. Homodimeric GFP and mCherry ligands showed that IFNAR2 but not IFNAR1 homodimers were sufficient to induce STAT1/2 signaling. Transcriptome analysis revealed that synthetic murine type I IFN signaling was highly comparable to IFNα4 signaling. Moreover, replication of vesicular stomatitis virus (VSV) in a cell culture-based viral infection model using MC57 cells was significantly inhibited after stimulation with synthetic ligands. Using intracellular deletion variants and point mutations, Y510 and Y335 in murine IFNAR2 were verified as unique phosphorylation sites for STAT1/2 activation, whereas the other tyrosine residues in IFNAR1 and IFNAR2 were not involved in STAT1/2 phosphorylation. Comparative analysis of synthetic human IFNARs supports this finding. In summary, our data showed that synthetic type I IFN signal transduction is originating from IFNAR2 rather than IFNAR1.https://www.frontiersin.org/articles/10.3389/fmicb.2022.947169/fullinterferonsyntheticcytokinevirusnanobody |
| spellingShingle | Nele Zoellner Noémi Coesfeld Frederik Henry De Vos Jennifer Denter Haifeng C. Xu Elena Zimmer Birgit Knebel Hadi Al-Hasani Sofie Mossner Philipp A. Lang Doreen M. Floss Jürgen Scheller Synthetic mimetics assigned a major role to IFNAR2 in type I interferon signaling Frontiers in Microbiology interferon synthetic cytokine virus nanobody |
| title | Synthetic mimetics assigned a major role to IFNAR2 in type I interferon signaling |
| title_full | Synthetic mimetics assigned a major role to IFNAR2 in type I interferon signaling |
| title_fullStr | Synthetic mimetics assigned a major role to IFNAR2 in type I interferon signaling |
| title_full_unstemmed | Synthetic mimetics assigned a major role to IFNAR2 in type I interferon signaling |
| title_short | Synthetic mimetics assigned a major role to IFNAR2 in type I interferon signaling |
| title_sort | synthetic mimetics assigned a major role to ifnar2 in type i interferon signaling |
| topic | interferon synthetic cytokine virus nanobody |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2022.947169/full |
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