title: Quantitative Proteomic Approach Identifies Vpr Binding Protein as Novel Host Factor Supporting Influenza A Virus Infections in Human Cells.
abstract: Influenza A virus (IAV) infections are a major cause for respiratory disease in humans, which affects all age groups and contributes substantially to global morbidity and mortality. IAV have a large natural host reservoir in avian species. However, many avian IAV strains lack adaptation to other hosts and hardly propagate in humans. While seasonal or pandemic IAV strains replicate efficiently in permissive human cells, many avian IAV cause abortive nonproductive infections in these hosts despite successful cell entry. However, the precise reasons for these differential outcomes are poorly defined. We hypothesized that the distinct course of an IAV infection with a given virus strain is determined by the differential interplay between specific host and viral factors. By using Spike-in SILAC mass spectrometry-based quantitative proteomics we characterized sets of cellular factors whose abundance is specifically up- or downregulated in the course of permissiveversusnonpermissive IAV infection, respectively. This approach allowed for the definition and quantitative comparison of about 3500 proteins in human lung epithelial cells in response to seasonal or low-pathogenic avian H3N2 IAV. Many identified proteins were similarly regulated by both virus strains, but also 16 candidates with distinct changes in permissiveversusnonpermissive infection were found. RNAi-mediated knockdown of these differentially regulated host factors identified Vpr binding protein (VprBP) as proviral host factor because its downregulation inhibited efficient propagation of seasonal IAV whereas overexpression increased viral replication of both seasonal and avian IAV. These results not only show that there are similar differences in the overall changes during permissive and nonpermissive influenza virus infections, but also provide a basis to evaluate VprBP as novel anti-IAV drug target.
authors.affiliation: From the ‡Unit 17 Influenza and other Respiratory Viruses", Robert Koch Institut, Seestr. 10, 13353 Berlin, Germany.
authors.forename: Anne
authors.initials: A
authors.lastname: Sadewasser
authors.affiliation: From the ‡Unit 17 Influenza and other Respiratory Viruses", Robert Koch Institut, Seestr. 10, 13353 Berlin, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Paki
authors.affiliation: §Max-Delbrück-Center for Molecular Medicine, Robert-Rössle-Str. 10, 13125 Berlin, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Eichelbaum
authors.affiliation: §Max-Delbrück-Center for Molecular Medicine, Robert-Rössle-Str. 10, 13125 Berlin, Germany.
authors.forename: Boris
authors.initials: B
authors.lastname: Bogdanow
authors.affiliation: From the ‡Unit 17 Influenza and other Respiratory Viruses", Robert Koch Institut, Seestr. 10, 13353 Berlin, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Saenger
authors.affiliation: From the ‡Unit 17 Influenza and other Respiratory Viruses", Robert Koch Institut, Seestr. 10, 13353 Berlin, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Budt
authors.affiliation: ¶Max Planck Institute for Infection Biology, Charitéplatz, 110117 Berlin, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Lesch
authors.affiliation: ‖Institute of Inorganic and Analytical Chemistry, Justus Liebig University, Heinrich-Buff-Ring 17, 35392 Giessen, Germany.
authors.forename: Klaus-Peter
authors.initials: KP
authors.lastname: Hinz
authors.affiliation: **Molecular Biophysics, Department of Biology, Humboldt-Universität zu Berlin, Invalidenstr. 43, 10115 Berlin, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Herrmann
authors.affiliation: ¶Max Planck Institute for Infection Biology, Charitéplatz, 110117 Berlin, Germany.
authors.forename: Thomas F
authors.initials: TF
authors.lastname: Meyer
authors.affiliation: ¶Max Planck Institute for Infection Biology, Charitéplatz, 110117 Berlin, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Karlas
authors.affiliation: §Max-Delbrück-Center for Molecular Medicine, Robert-Rössle-Str. 10, 13125 Berlin, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Selbach
authors.affiliation: From the ‡Unit 17 Influenza and other Respiratory Viruses", Robert Koch Institut, Seestr. 10, 13353 Berlin, Germany; WolffT@RKI.de.
authors.forename: Thorsten
authors.initials: T
authors.lastname: Wolff
mesh_terms: D000072283:A549 Cells; D002352:Carrier Proteins / Q000378:metabolism*; D016000:Cluster Analysis; D004705:Endocytosis; D004847:Epithelial Cells / Q000378:metabolism* / Q000473:pathology / Q000821:virology*; D057809:HEK293 Cells; D006801:Humans; D053122:Influenza A Virus, H3N2 Subtype / Q000502:physiology*; D007251:Influenza, Human / Q000821:virology*; D007553:Isotope Labeling; D008168:Lung / Q000473:pathology; D013058:Mass Spectrometry; D017346:Protein Serine-Threonine Kinases; D020543:Proteome / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D034741:RNA, Small Interfering / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases; D014779:Virus Replication
chemical_list: D002352:Carrier Proteins; D020543:Proteome; D034741:RNA, Small Interfering; D044767:Ubiquitin-Protein Ligases; C530533:DCAF1 protein, human; D017346:Protein Serine-Threonine Kinases