ERA NanoSci - Nanocommunication: How virus particles convince cells to let them inside

• Antragstellerinnen / Antragsteller: Professor Dr. Kay Grünewald; Professorin Dr. Beate Sodeik
• Fachliche Zuordnung: Strukturbiologie ; Theoretische Chemie: Elektronenstruktur, Dynamik, Simulation
• Förderung: Förderung von 2009 bis 2015
• Projektkennung: Deutsche Forschungsgemeinschaft (DFG) - Projektnummer 118558436

• Erstellungsjahr: 2016

Zusammenfassung der Projektergebnisse

The ERAnet nanoSciE+ network brought together complementary expertise of four groups from four countries (Netherlands, Israel, Germany, UK) to collaboratively target key aspects in the ‘nano-communication’ that underlies virus entry. Eukaryotic viruses replicating in the nucleus must pass two host barriers, the plasma membranes and the nuclear envelopes, to deliver their genomes to a compartment allowing viral transcription and replication, and to initiate an infection. Viral nanomachines have evolved to efficiently interface with and overcome these host barriers. The network activities were organised in five workpackages to understand this molecular interfacing better by using our combined expertise. The DFG-funded groups of Beate Sodeik and Kay Grünewald contributed to all workpackages. The synergistic interaction in the network allowed for a broader methodological approach and to establish novel investigation systems. A highlight example is the work that led to the Traffic publication (by Israeli plus the two German groups). It describes a newly established in vitro system to study the interactions of herpesvirus capsids with nuclear pore complexes and revealed a first set of (co)-factors required, opening the way to better understand the physical, structural and biochemical principles underlying the functional coupling that induces genome uncoating into the nucleus (second virus entry barrier). Another highlight outcome is the establishment of a successful experimental system for displaying full-length membrane proteins on extracellular vesicles for structural studies. This paved the way to our just published first characterisation of the hitherto unknown pre-fusion structure of the conserved herpesvirus key fusion machinery element, glycoprotein B (PNAS, 2016). There is clear excitement about this structure (coverage in Science) as making this metastable and for interventions critical state tractable will surely induce novel research hypothesis even beyond the field of herpesvirus membrane fusion (first virus entry barrier). Altogether, both biological and methodological advances have been achieved. A couple of them has significance for future developments in the field by paving the way as experimental setup (in vitro HSV1-NPC interaction) or their insights (gB pre-fusion structure study). Within Germany, the DFG-funded groups plan to continue their successful collaboration. Favoured by the immanent return of the Grünewald group to Germany in which the MHH (home of the Sodeik group) is also a partner, the groups are currently involved in planning a joint graduate college between the two sites.

Projektbezogene Publikationen (Auswahl)

• (2011). Cryo Electron Tomography of Herpes Simplex Virus during Axonal Transport and Secondary Envelopment in Primary Neurons. PLoS Pathogens 7:e1002406
  Ibiricu I, Huiskonen JT, Döhner K, Bradke F, Sodeik B, Grünewald K
  
• (2011). Uncoupling uncoating of Herpes Simplex Virus genomes from their nuclear import and gene expression. Journal of Virology, 85:4271-4283 (Cover Image and Spotlight)
  Rode R, K Döhner, A Binz, M Glass, T Strive, R Bauerfeind & B Sodeik
  
• (2013). Characterization of herpes simplex virus type 1 L-particle assembly and egress in hippocampal neurones by electron cryo-tomography. Cellular Microbiol. 15:285-291
  Ibiricu I, UE Maurer, K Grünewald
  
• (2013). Structure of herpesvirus fusion glycoprotein B- bilayer complex revealing the protein-membrane and lateral protein-protein interaction. Structure 21:1396-1405
  Maurer UE, T Zeev-Ben-Mordehai, AP Pandurangan, TM Cairns, BP Hannah, JC Whitbeck, RJ Eisenberg, GH Cohen, M Topf, JT Huiskonen, K Grünewald
  
• (2014), A cool hybrid approach to the herpesvirus ‘life’ cycle Curr. Opin. Virol. 5: 42–49
  Zeev-Ben-Mordehai T, C Hagen, K Grünewald
  
• (2014). A precipitation-based assay to analyze interactions of viral particles with cytosolic host factors. Methods in Molecular Biology 1144:191-208
  Radtke K, F Anderson & B Sodeik
  
• (2014). The full-length cell-cell fusogen EFF-1 is monomeric and upright to the membrane. Nature Communications 5: 3912-3920
  Zeev-Ben-Mordehai T, D Vasishtan, CA Siebert, K Grünewald
  
• Specific targeting of Herpes Simplex Virus Capsids to Nuclear Pores in a cell-free nuclear reconstitution system. Traffic 15:1266-1281 (Cover Image)
  Anderson F, A Savulescu, K Rudolph, J Schipke, I Cohen, I Ibiricu, A Rotem, K Grünewald, B Sodeik & A Harel
  
• (2015). The amphipatic helix of adenovirus capsid protein VI contributes to penton release and post entry sorting. J. Virol. 89: 2121-35
  Martinez R, P Schellenberger, D Vasishtan, C Aknin, S Austin, D Dacheux, F Rayne, CA Siebert, Ruzsics, Z., K Grünewald, H Wodrich
  
• (2016). Two distinct trimeric conformations of natively membrane-anchored full-length Herpes simplex virus 1 glycoprotein B. PNAS
  Zeev-Ben-Mordehai T, D Vasishtan, A Hernández Durán, B Vollmer, P White, AP Pandurangan, CA Siebert, M Topf, K Grünewald