title: New host factors important for respiratory syncytial virus (RSV) replication revealed by a novel microfluidics screen for interactors of matrix (M) protein.
abstract: Although human respiratory syncytial virus (RSV) is the most common cause of bronchiolitis and pneumonia in infants and elderly worldwide, there is no licensed RSV vaccine or effective drug treatment available. The RSV Matrix protein plays key roles in virus life cycle, being found in the nucleus early in infection in a transcriptional inhibitory role, and later localizing in viral inclusion bodies before coordinating viral assembly and budding at the plasma membrane. In this study, we used a novel, high throughput microfluidics platform and custom human open reading frame library to identify novel host cell binding partners of RSV matrix. Novel interactors identified included proteins involved in host transcription regulation, the innate immunity response, cytoskeletal regulation, membrane remodeling, and cellular trafficking. A number of these interactions were confirmed by immunoprecipitation and cellular colocalization approaches. Importantly, the physiological significance of matrix interaction with the actin-binding protein cofilin 1, caveolae protein Caveolin 2, and the zinc finger protein ZNF502 was confirmed. siRNA knockdown of the host protein levels resulted in reduced RSV virus production in infected cells. These results have important implications for future antiviral strategies aimed at targets of RSV matrix in the host cell.
authors.affiliation: From the ‡Nanotechnology Institute, Mina and Evrard Goodman Faculty of Life Sciences, Bar Ilan University, Ramat Gan, Israel;
authors.forename: Sarit
authors.initials: S
authors.lastname: Kipper
authors.affiliation: §Section of Virology, Faculty of Medicine, Imperial College London, London, UK;
authors.forename: Samar
authors.initials: S
authors.lastname: Hamad
authors.affiliation: ¶Department of Biochemistry and Molecular Biology, Monash University, Melbourne, Australia;
authors.forename: Leon
authors.initials: L
authors.lastname: Caly
authors.affiliation: From the ‡Nanotechnology Institute, Mina and Evrard Goodman Faculty of Life Sciences, Bar Ilan University, Ramat Gan, Israel;
authors.forename: Dorit
authors.initials: D
authors.lastname: Avrahami
authors.affiliation: ‖Department of Cell Research and Immunology, Tel Aviv University, Israel.
authors.forename: Eran
authors.initials: E
authors.lastname: Bacharach
authors.affiliation: ¶Department of Biochemistry and Molecular Biology, Monash University, Melbourne, Australia;
authors.forename: David A
authors.initials: DA
authors.lastname: Jans
authors.affiliation: From the ‡Nanotechnology Institute, Mina and Evrard Goodman Faculty of Life Sciences, Bar Ilan University, Ramat Gan, Israel; Doron.Gerber@biu.ac.il m.bajorek@imperial.ac.uk.
authors.forename: Doron
authors.initials: D
authors.lastname: Gerber
authors.affiliation: §Section of Virology, Faculty of Medicine, Imperial College London, London, UK; Doron.Gerber@biu.ac.il m.bajorek@imperial.ac.uk.
authors.forename: Monika
authors.initials: M
authors.lastname: Bajorek
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D051243:Caveolin 2 / Q000378:metabolism*; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002522:Chlorocebus aethiops; D051338:Cofilin 1 / Q000378:metabolism*; D015723:Gene Library; D057809:HEK293 Cells; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D009687:Nuclear Proteins / Q000378:metabolism*; D016366:Open Reading Frames; D012136:Respiratory Syncytial Viruses / Q000502:physiology*; D014709:Vero Cells; D014763:Viral Matrix Proteins / Q000378:metabolism*; D014779:Virus Replication
chemical_list: C495544:CAV2 protein, human; C495810:CFL1 protein, human; D002352:Carrier Proteins; D051243:Caveolin 2; D051338:Cofilin 1; D009687:Nuclear Proteins; D014763:Viral Matrix Proteins; C000603037:ZNF502 protein, human