1. Proteomic Analysis Uncovers Measles Virus Protein C Interaction With p65-iASPP Protein Complex
- Author
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Meignié, Alice, Combredet, Chantal, Santolini, Marc, Kovács, István, Douché, Thibaut, Gianetto, Quentin Giai, Eun, Hyeju, Matondo, Mariette, Jacob, Yves, Grailhe, Regis, Tangy, Frédéric, Komarova, Anastassia, Génomique virale et vaccination, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Université Paris Diderot - Paris 7 (UPD7), Centre de Recherche Interdisciplinaire / Center for Research and Interdisciplinarity [Paris, France] (CRI), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP), Northeastern University [Boston], Northwestern University [Evanston], Central European University [Budapest, Hongrie] (CEU), Plateforme de Protéomique / Proteomics platform, Spectrométrie de Masse pour la Biologie – Mass Spectrometry for Biology (UTechS MSBio), Institut Pasteur [Paris]-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris]-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Hub Bioinformatique et Biostatistique - Bioinformatics and Biostatistics HUB, Institut Pasteur Korea - Institut Pasteur de Corée, Réseau International des Instituts Pasteur (RIIP), Génétique Moléculaire des Virus à ARN - Molecular Genetics of RNA Viruses (GMV-ARN (UMR_3569 / U-Pasteur_2)), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), This work was supported by ANR 16 CE18 0016 01 Tangy ONCOMEVAX, the Institut Pasteur, and CNRS., We thank Dr Kaoru Takeuchi and Dr Chantal Rabourdin-Combe for anti-C rAb polyclonal antibody and anti-N monoclonal antibody, respectively. We thank Dr.Hervé Bourhy from Lyssavirus Epidemiology and Neuropathology Laboratory of Institut Pasteur for providing us pYFP-p65 plasmid that contains EYFP reporter gene fused to p65. We thank all members of the Viral Genomics and Vaccination Unit and Laboratory of Molecular Genetics of RNA Viruses for their critical discussion, Dr Atousa Arbabian for technical support with experiments, Dr Dmitry Trubetskoy for the help with data representation, and Dr Aleksandr Barinov and Dr Caroline Demeret for critical reading., ANR-16-CE18-0016,OncoMeVax,Un virus de la rougeole modifié pour traiter le cancer(2016), Matondo, Mariette, Un virus de la rougeole modifié pour traiter le cancer - - OncoMeVax2016 - ANR-16-CE18-0016 - AAPG2016 - VALID, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Institut Pasteur [Paris] (IP)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris] (IP)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité)
- Subjects
Proteomics ,RIG-I, retinoic acid–inducible gene-I ,FDR, false discovery rate ,NF-kB, nuclear factor-kappa B ,PPI, protein–protein interaction ,virus-host protein-protein interactions ,virulence factors ,SOD1, superoxide dismutase 1 ,Viral Nonstructural Proteins ,Virus Replication ,Cell Line ,PKR, protein kinase R ,Chlorocebus aethiops ,BRET, bioluminescence resonance energy Transfer ,[SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology ,Animals ,Humans ,AGC, automatic Gain Control ,[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology ,IFN, interferon ,LCC, largest connected component ,Protein Interaction Maps ,NLR, normalized luminescence ratio ,innate immunity ,MOI, multiplicity of infection ,FA, formic acid ,DCA, dichloroacetic acid ,Cell Death ,Research ,Y2H, yeast two-hybrid ,Intracellular Signaling Peptides and Proteins ,Transcription Factor RelA ,ACN, acetonitrile ,MV, measles virus ,Repressor Proteins ,ATU, additional transcription unit ,Measles virus ,Host-Pathogen Interactions ,DI, defective interfering ,PCA, protein complementation assay ,Tumor Suppressor Protein p53 ,Nluc, nanoluciferase ,Measles - Abstract
Viruses manipulate the central machineries of host cells to their advantage. They prevent host cell antiviral responses to create a favorable environment for their survival and propagation. Measles virus (MV) encodes two nonstructural proteins MV-V and MV-C known to counteract the host interferon response and to regulate cell death pathways. Several molecular mechanisms underlining MV-V regulation of innate immunity and cell death pathways have been proposed, whereas MV-C host-interacting proteins are less studied. We suggest that some cellular factors that are controlled by MV-C protein during viral replication could be components of innate immunity and the cell death pathways. To determine which host factors are targeted by MV-C, we captured both direct and indirect host-interacting proteins of MV-C protein. For this, we used a strategy based on recombinant viruses expressing tagged viral proteins followed by affinity purification and a bottom-up mass spectrometry analysis. From the list of host proteins specifically interacting with MV-C protein in different cell lines, we selected the host targets that belong to immunity and cell death pathways for further validation. Direct protein interaction partners of MV-C were determined by applying protein complementation assay and the bioluminescence resonance energy transfer approach. As a result, we found that MV-C protein specifically interacts with p65–iASPP protein complex that controls both cell death and innate immunity pathways and evaluated the significance of these host factors on virus replication., Graphical Abstract, Highlights • Measles virus controls immune response and cell death pathways to achieve replication. • Host proteins interaction network with measles virulence factor C protein. • Cellular p65–iASPP complex is targeted by measles virus C protein., In Brief Control of measles virus infection and measles-based oncolytic therapy are possible, thanks to the existence of a safe and efficient live attenuated vaccine. Molecular mechanisms that make this vaccine to be so efficient are yet to be determined. We show that the measles C protein is responsible for the establishment of complex networks of interactions with the host cell. We suggest that the C protein binding to the p65–iASPP protein complex controls the host cell death and innate immunity pathways.
- Published
- 2020