Identification of a short sequence motif in the influenza A virus pathogenicity factor PB1-F2 required for inhibition of human NLRP3

Article Article scientifique Article Identification of a short sequence motif in the influenza A virus pathogenicity factor PB1-F2 required for inhibition of human NLRP3 eng 2024-04-03 2024-04-03 author Silva Filo Department of Microbiology and Molecular Medicine, Medical Faculty, University of Geneva, Geneva, Switzerland 23504 Département de microbiologie et médecine moléculaire (MIMOL) author Boal P. T. De Carvalho Inès Department of Microbiology and Molecular Medicine, Medical Faculty, University of Geneva, Geneva, Switzerland 828310 author Williams Nathalia 0000-0003-2279-5302 Department of Microbiology and Molecular Medicine, Medical Faculty, University of Geneva, Geneva, Switzerland 942117 Département de microbiologie et médecine moléculaire (MIMOL) author Chabert-Ben Cherifa Mehdi Department of Microbiology and Molecular Medicine, Medical Faculty, University of Geneva, Geneva, Switzerland 410694 Département de microbiologie et médecine moléculaire (MIMOL) author Niu Chengyue 0000-0001-8607-3419 Department of Microbiology and Molecular Medicine, Medical Faculty, University of Geneva, Geneva, Switzerland 805693 author Hedhili Dalila Sabrine Department of Microbiology and Molecular Medicine, Medical Faculty, University of Geneva, Geneva, Switzerland 926683 author Choltus Helena Department of Microbiology and Molecular Medicine, Medical Faculty, University of Geneva, Geneva, Switzerland 984092 Département de microbiologie et médecine moléculaire (MIMOL) author Liaudet Nicolas 0000-0002-0749-5857 Bioimaging Core Facility, Medical Faculty, University of Geneva, Geneva, Switzerland 862067 Services communs de la faculté de médecine (SCMU) author Gaia Nadia Genomic Research Laboratory, Division of Infectious Diseases, Department of Medicine, University Hospitals and University of Geneva, Geneva, Switzerland 800637 author Karenovics Wolfram Thoracic Surgery, Geneva University Hospitals, Geneva, Switzerland 501317 author Francois Patrice 0000-0002-0540-750X Department of Microbiology and Molecular Medicine, Medical Faculty, University of Geneva, Geneva, Switzerland 28911 Département de médecine (DEMED) author Schmolke Mirco 0000-0002-2491-3029 Department of Microbiology and Molecular Medicine, Medical Faculty, University of Geneva, Geneva, Switzerland 821513 Département de microbiologie et médecine moléculaire (MIMOL) e0041124 10.1128/jvi.00411-24 38567952 0022-538X https://journals.asm.org/doi/10.1128/jvi.00411-24 <p>Influenza A virus infection activates the NLRP3 inflammasome, a multiprotein signaling complex responsible for the proteolytic activation and release of the proinflammatory cytokine IL-1β from monocytes and macrophages. Some influenza A virus (IAV) strains encode a short 90-amino acid peptide (PB1-F2) on an alternative open reading frame of segment 2, with immunomodulatory activity. We recently demonstrated that contemporary IAV PB1-F2 inhibits the activation of NLRP3, potentially by NEK7-dependent activation. PB1-F2 binds to NLRP3 with its C-terminal 50 amino acids, but the exact binding motif was unknown. On the NLRP3 side, the interface is formed through the leucine-rich-repeat (LRR) domain, potentially in conjunction with the pyrin domain. Here, we took advantage of PB1-F2 sequences from IAV strains with either weak or strong NLRP3 interaction. Sequence comparison and structure prediction using Alphafold2 identified a short four amino acid sequence motif (TQGS) in PB1-F2 that defines NLRP3-LRR binding. Conversion of this motif to that of the non-binding PB1-F2 suffices to lose inhibition of NLRP3 dependent IL-1β release. The TQGS motif further alters the subcellular localization of PB1-F2 and its colocalization with NLRP3 LRR and pyrin domain. Structural predictions suggest the establishment of additional hydrogen bonds between the C-terminus of PB1-F2 and the LRR domain of NLRP3, with two hydrogen bonds connecting to threonine and glutamine of the TQGS motif. Phylogenetic data show that the identified NLRP3 interaction motif in PB1-F2 is widely conserved among recent IAV-infecting humans. Our data explain at a molecular level the specificity of NLRP3 inhibition by influenza A virus.</p> IMPORTANCE <p>Influenza A virus infection is accompanied by a strong inflammatory response and high fever. The human immune system facilitates the swift clearance of the virus with this response. An essential signal protein in the proinflammatory host response is IL-1b. It is released from inflammatory macrophages, and its production and secretion depend on the function of NLRP3. We had previously shown that influenza A virus blocks NLRP3 activation by the expression of a viral inhibitor, PB1-F2. Here, we demonstrate how this short peptide binds to NLRP3 and provide evidence that a four amino acid stretch in PB1-F2 is necessary and sufficient to mediate this binding. Our data identify a new virus-host interface required to block one signaling path of the innate host response against influenza A virus.</p> IL-1beta NLRP3 PB1-F2 Inflammasome Inflammation Influenza Macrophage Protein interactions Dewey 616 Dewey 617 Swiss National Science Foundation Identification and characterization of host factors affecting Inter-and intra-species spreading of influenza A viruses 212216 CH Projects Swiss National Science Foundation Interaction of Influenza A virus with pathogenic and commensal bacteria 155949 CH Projects 54 Département de microbiologie et médecine moléculaire 21B 57 Département de chirurgie 222 false 912 Modulation de la flore microbienne dans un hôte infecté par un virus 947 338 Génomique et microbiote 604 902 Cancer du poumon et mésothéliome pleural malin 944 Journal of virology d5154603-eac6-485c-82b8-ab37f70e6785 Article (Published version) Public Article-scientifique-from-crossref.pdf 1603091 application/pdf Restricted 2024-10-04 bfe5998e6af8f47a5fbdde1713a011a5bb9ab568d824d4de873d0330fb632f35 39277561-2473-4412-b44b-a1a1a15523f1 Supplemental data Public jvi.00411-24-s0001.eps 1488310 application/postscript b438b3fe1ff0bc13d0136a0258f1b6936ea08f9cd98577f3aed5088230580713 13788c7c-259a-45dd-8966-6fb428f3a430 Supplemental data Public jvi.00411-24-s0002.pdf 51352 application/pdf 82db394dfae31077d89bbc1888668a45dcbd9cd1e063ecf847cafbe6e916e785 cade5cd7-5485-478f-9278-2100220b6329 Supplemental data Public jvi.00411-24-s0003(1).eps 251666 application/postscript 8fd647c162487a72d0722383f19ff2344b06f665c7f9ca047107e4dd96e10172 54afe8c3-3aaf-4842-a1a5-130060e46d6b Supplemental data Public jvi.00411-24-s0004.eps 1171019 application/postscript 69742b537d02b201517e081541bf8d63117b7e7044871952ed97ebd1123af273 d28eb60b-ef84-42aa-af6f-b7d63b9e02dc Supplemental data Public jvi.00411-24-s0005.eps 188233 application/postscript 578086efeedb1cd6f89f23f72622746f5139c0cfc9da0f23ca5a6c70bd9ce361 080fd714-5f79-4b89-883a-7528d31339c6 Supplemental data Public jvi.00411-24-s0006.eps 6494008 application/postscript 809d95a49138c79f701e0911a4aa7763fc5912a14ffb1c31cbd219758c0e830d 84c24cf8-72c9-462c-8309-d434208aca8d Supplemental data Public jvi.00411-24-s0007.eps 260419 application/postscript 665a25b5258ef5550fe02ec5c4f3ddc4a1bd87dd77bc7e6dab7fd05871c0108f 02ead61e-5bd8-48fc-a135-797461a518bb Supplemental data Public jvi.00411-24-s0008.docx 120946 application/vnd.openxmlformats-officedocument.wordprocessingml.document fa03eb2770fdb0e85864dd789df7a6a73424b26d8093ea8959d016af1a8a2e32 7b24d3c7-aee4-46a0-a36d-3e2411cdb71e Supplemental data Public jvi.00411-24-s0009.pdf 51008 application/pdf 34b7c8025e84c685f5c07c1bad5c9c0bba68482ebf99a32c69cfb9ea3b6f47ce 210207c6-c007-40b8-b8e2-32f0aa67b0e2 Supplemental data Public jvi.00411-24-s0010.pdf 43712 application/pdf 694aa8c4bafe409cdca68918288c8bea20db1cecb76217edc250df273be768f0 openAccess Faculté de médecine / Section de médecine fondamentale / Département de microbiologie et médecine moléculaire Faculté de médecine / Section de médecine clinique / Département de chirurgie

Title	Published in	Year	Views	Downloads

Identification of a short sequence motif in the influenza A virus pathogenicity factor PB1-F2 required for inhibition of human NLRP3	Journal of virology	2024	259	252
Defining the molecular interface of Influenza-A-virus pathogenicity factor PB1-F2 with human inflammasome NLRP3		2021	89	6

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