Influenza virus protein PB1-F2 interacts with CALCOCO2 (NDP52) to modulate innate immune response

PB1-F2 is a viral protein encoded by influenza A viruses (IAVs). PB1-F2 is implicated in virulence by triggering immune cell apoptosis and enhancing inflammation. To obtain an insight into the molecular mechanisms of PB1-F2-mediated virulence, we used the yeast two-hybrid approach to find new PB1-F2 cellular interactors. This allowed us to identify calcium-binding and coiled-coil domain 2 (CALCOCO2, also known as NDP52) as a binding partner of PB1-F2. Binding of PB1-F2 to CALCOCO2 was confirmed by pull-down. Surface plasmon resonance binding experiments enabled us to estimate the dissociation constant (Kd) of the two partners to be around 20 nM. Using bioinformatics tools, we designed a CALCOCO2 interaction map based on previous knowledge and showed a strong connection between this protein and the type I interferon production pathways and the I-\kappaB kinase/NF-\kappaB signalling pathway. NF-\kappaB reporter assays in which CALCOCO2, MAVS and PB1-F2 were co-expressed showed a cooperation of these three proteins to increase the inflammatory response. By contrast, PB1-F2 inhibits the TBK1-dependent activation of an ISRE reporter plasmid. We also demonstrated that the signal transducer TRAF6 is implicated in the enhancement of NF-\kappaB activity mediated by PB1-F2/CALCOCO2 binding. Altogether, this report provides evidence of an interaction link between PB1-F2 and human proteins, and allows a better understanding of the involvement of PB1-F2 in the pathologic process mediated by IAV.

Mots clés

Protein Binding Protein Interaction Mapping Surface Plasmon Resonance Computational Biology Host-Pathogen Interactions Humans Immunity Innate Influenza A virus Kinetics Nuclear Proteins Two-Hybrid System Techniques Viral Proteins Virulence Factors PB1-F2 CALCOCO2 calcium-binding and coiled-coil domain 2 IAV influenza A virus ISRE IFN-stimulated response element MAVS mitochondrial antiviral signalling protein qRT-PCR quantitative reverse transcription PCR SPR surface plasmon resonance siRNA small interfering RNA TRAF6 TNF receptor-associated factor 6 TRAF6 DN dominant-negative form of TRAF6 TRAF6 FL full-length TRAF6 Y2H yeast two-hybrid system RU resonance unit influenza virus innate immune response

Domaines

Immunologie Bactériologie Virologie

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https://hal.science/hal-01911416

Soumis le : vendredi 2 novembre 2018-17:39:51

Dernière modification le : mercredi 3 juillet 2024-09:00:06

Dates et versions

hal-01911416 , version 1 (02-11-2018)

Identifiants

HAL Id : hal-01911416 , version 1
DOI : 10.1099/jgv.0.000782
PRODINRA : 413121
PUBMED : 28613140
WOS : 000410020900009

Citer

Olivier Leymarie, Léa Meyer, Lionel Tafforeau, Vincent Lotteau, Bruno da Costa, et al.. Influenza virus protein PB1-F2 interacts with CALCOCO2 (NDP52) to modulate innate immune response. Journal of General Virology, 2017, 98 (6), pp.1196-1208. ⟨10.1099/jgv.0.000782⟩. ⟨hal-01911416⟩

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