Novel function of Bleutongue Virus NS3 Protein in Regulation of the MAPK/ERK Signaling Pathway

Cindy Kundlacz, Marie Pourcelot, Aurore Fablet, Rayane Amaral Da Silva Moraes, Thibaut Leger, Bastien Morlet, Cyril Viarouge, C. Sailleau, Mathilde Turpaud, Axel Gorlier, Emmanuel Breard, S. Lecollinet, P.A. van Rijn, Stephan Zientara, Damien Vitour, Gregory Caignard

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Bluetongue virus (BTV) is an arbovirus transmitted by blood-feeding midges to a wide range of wild and domestic ruminants. In this report, we showed that BTV, through its nonstructural protein NS3 (BTV-NS3), is able to activate the mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway, as assessed by phosphorylation levels of ERK1/2 and the translation initiation factor eukaryotic translation initiation factor 4E (eIF4E). By combining immunoprecipitation of BTV-NS3 and mass spectrometry analysis from both BTV-infected and NS3-transfected cells, we identified the serine/threonine-protein kinase B-Raf (BRAF), a crucial player in the MAPK/ERK pathway, as a new cellular interactor of BTV-NS3. BRAF silencing led to a significant decrease in the MAPK/ERK activation by BTV, supporting a model wherein BTV-NS3 interacts with BRAF to activate this signaling cascade. This positive regulation acts independently of the role of BTV-NS3 in counteracting the induction of the alpha/beta interferon response. Furthermore, the intrinsic ability of BTV-NS3 to bind BRAF and activate the MAPK/ERK pathway is conserved throughout multiple serotypes/strains but appears to be specific to BTV compared to other members of Orbivirus genus. Inhibition of MAPK/ERK pathway with U0126 reduced viral titers, suggesting that BTV manipulates this pathway for its own replication. Altogether, our data provide molecular mechanisms that unravel a new essential function of NS3 during BTV infection.
LanguageEnglish
Article numbere00336-19
Number of pages17
JournalJournal of Virology
Volume93
Issue number16
DOIs
Publication statusPublished - Aug 2019

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Bluetongue virus
Extracellular Signal-Regulated MAP Kinases
Mitogen-Activated Protein Kinases
mitogen-activated protein kinase
Viruses
viruses
Proteins
proteins
translation (genetics)
Orbivirus
Eukaryotic Initiation Factor-4E
Eukaryotic Initiation Factors
Arboviruses
interferon-beta
Proto-Oncogene Proteins c-akt
arboviruses
Interferon-beta
Protein-Serine-Threonine Kinases
Ruminants
midges

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Kundlacz, C., Pourcelot, M., Fablet, A., Amaral Da Silva Moraes, R., Leger, T., Morlet, B., ... Caignard, G. (2019). Novel function of Bleutongue Virus NS3 Protein in Regulation of the MAPK/ERK Signaling Pathway. Journal of Virology, 93(16), [e00336-19]. https://doi.org/10.1128/JVI.00336-19
Kundlacz, Cindy ; Pourcelot, Marie ; Fablet, Aurore ; Amaral Da Silva Moraes, Rayane ; Leger, Thibaut ; Morlet, Bastien ; Viarouge, Cyril ; Sailleau, C. ; Turpaud, Mathilde ; Gorlier, Axel ; Breard, Emmanuel ; Lecollinet, S. ; van Rijn, P.A. ; Zientara, Stephan ; Vitour, Damien ; Caignard, Gregory. / Novel function of Bleutongue Virus NS3 Protein in Regulation of the MAPK/ERK Signaling Pathway. In: Journal of Virology. 2019 ; Vol. 93, No. 16.
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title = "Novel function of Bleutongue Virus NS3 Protein in Regulation of the MAPK/ERK Signaling Pathway",
abstract = "Bluetongue virus (BTV) is an arbovirus transmitted by blood-feeding midges to a wide range of wild and domestic ruminants. In this report, we showed that BTV, through its nonstructural protein NS3 (BTV-NS3), is able to activate the mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway, as assessed by phosphorylation levels of ERK1/2 and the translation initiation factor eukaryotic translation initiation factor 4E (eIF4E). By combining immunoprecipitation of BTV-NS3 and mass spectrometry analysis from both BTV-infected and NS3-transfected cells, we identified the serine/threonine-protein kinase B-Raf (BRAF), a crucial player in the MAPK/ERK pathway, as a new cellular interactor of BTV-NS3. BRAF silencing led to a significant decrease in the MAPK/ERK activation by BTV, supporting a model wherein BTV-NS3 interacts with BRAF to activate this signaling cascade. This positive regulation acts independently of the role of BTV-NS3 in counteracting the induction of the alpha/beta interferon response. Furthermore, the intrinsic ability of BTV-NS3 to bind BRAF and activate the MAPK/ERK pathway is conserved throughout multiple serotypes/strains but appears to be specific to BTV compared to other members of Orbivirus genus. Inhibition of MAPK/ERK pathway with U0126 reduced viral titers, suggesting that BTV manipulates this pathway for its own replication. Altogether, our data provide molecular mechanisms that unravel a new essential function of NS3 during BTV infection.",
author = "Cindy Kundlacz and Marie Pourcelot and Aurore Fablet and {Amaral Da Silva Moraes}, Rayane and Thibaut Leger and Bastien Morlet and Cyril Viarouge and C. Sailleau and Mathilde Turpaud and Axel Gorlier and Emmanuel Breard and S. Lecollinet and {van Rijn}, P.A. and Stephan Zientara and Damien Vitour and Gregory Caignard",
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Kundlacz, C, Pourcelot, M, Fablet, A, Amaral Da Silva Moraes, R, Leger, T, Morlet, B, Viarouge, C, Sailleau, C, Turpaud, M, Gorlier, A, Breard, E, Lecollinet, S, van Rijn, PA, Zientara, S, Vitour, D & Caignard, G 2019, 'Novel function of Bleutongue Virus NS3 Protein in Regulation of the MAPK/ERK Signaling Pathway', Journal of Virology, vol. 93, no. 16, e00336-19. https://doi.org/10.1128/JVI.00336-19

Novel function of Bleutongue Virus NS3 Protein in Regulation of the MAPK/ERK Signaling Pathway. / Kundlacz, Cindy; Pourcelot, Marie; Fablet, Aurore; Amaral Da Silva Moraes, Rayane; Leger, Thibaut; Morlet, Bastien; Viarouge, Cyril; Sailleau, C.; Turpaud, Mathilde; Gorlier, Axel; Breard, Emmanuel; Lecollinet, S.; van Rijn, P.A.; Zientara, Stephan; Vitour, Damien; Caignard, Gregory.

In: Journal of Virology, Vol. 93, No. 16, e00336-19, 08.2019.

Research output: Contribution to journalArticleAcademicpeer-review

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AU - Kundlacz, Cindy

AU - Pourcelot, Marie

AU - Fablet, Aurore

AU - Amaral Da Silva Moraes, Rayane

AU - Leger, Thibaut

AU - Morlet, Bastien

AU - Viarouge, Cyril

AU - Sailleau, C.

AU - Turpaud, Mathilde

AU - Gorlier, Axel

AU - Breard, Emmanuel

AU - Lecollinet, S.

AU - van Rijn, P.A.

AU - Zientara, Stephan

AU - Vitour, Damien

AU - Caignard, Gregory

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N2 - Bluetongue virus (BTV) is an arbovirus transmitted by blood-feeding midges to a wide range of wild and domestic ruminants. In this report, we showed that BTV, through its nonstructural protein NS3 (BTV-NS3), is able to activate the mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway, as assessed by phosphorylation levels of ERK1/2 and the translation initiation factor eukaryotic translation initiation factor 4E (eIF4E). By combining immunoprecipitation of BTV-NS3 and mass spectrometry analysis from both BTV-infected and NS3-transfected cells, we identified the serine/threonine-protein kinase B-Raf (BRAF), a crucial player in the MAPK/ERK pathway, as a new cellular interactor of BTV-NS3. BRAF silencing led to a significant decrease in the MAPK/ERK activation by BTV, supporting a model wherein BTV-NS3 interacts with BRAF to activate this signaling cascade. This positive regulation acts independently of the role of BTV-NS3 in counteracting the induction of the alpha/beta interferon response. Furthermore, the intrinsic ability of BTV-NS3 to bind BRAF and activate the MAPK/ERK pathway is conserved throughout multiple serotypes/strains but appears to be specific to BTV compared to other members of Orbivirus genus. Inhibition of MAPK/ERK pathway with U0126 reduced viral titers, suggesting that BTV manipulates this pathway for its own replication. Altogether, our data provide molecular mechanisms that unravel a new essential function of NS3 during BTV infection.

AB - Bluetongue virus (BTV) is an arbovirus transmitted by blood-feeding midges to a wide range of wild and domestic ruminants. In this report, we showed that BTV, through its nonstructural protein NS3 (BTV-NS3), is able to activate the mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway, as assessed by phosphorylation levels of ERK1/2 and the translation initiation factor eukaryotic translation initiation factor 4E (eIF4E). By combining immunoprecipitation of BTV-NS3 and mass spectrometry analysis from both BTV-infected and NS3-transfected cells, we identified the serine/threonine-protein kinase B-Raf (BRAF), a crucial player in the MAPK/ERK pathway, as a new cellular interactor of BTV-NS3. BRAF silencing led to a significant decrease in the MAPK/ERK activation by BTV, supporting a model wherein BTV-NS3 interacts with BRAF to activate this signaling cascade. This positive regulation acts independently of the role of BTV-NS3 in counteracting the induction of the alpha/beta interferon response. Furthermore, the intrinsic ability of BTV-NS3 to bind BRAF and activate the MAPK/ERK pathway is conserved throughout multiple serotypes/strains but appears to be specific to BTV compared to other members of Orbivirus genus. Inhibition of MAPK/ERK pathway with U0126 reduced viral titers, suggesting that BTV manipulates this pathway for its own replication. Altogether, our data provide molecular mechanisms that unravel a new essential function of NS3 during BTV infection.

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Kundlacz C, Pourcelot M, Fablet A, Amaral Da Silva Moraes R, Leger T, Morlet B et al. Novel function of Bleutongue Virus NS3 Protein in Regulation of the MAPK/ERK Signaling Pathway. Journal of Virology. 2019 Aug;93(16). e00336-19. https://doi.org/10.1128/JVI.00336-19