MA/MSc, PhD Internship for EUGLOH program

Keywords: Vaccinia virus assembly, Cellular membrane remodelling, Protein expression, Protein-lipid interactions, Cell-free expression system

Internship Duration: 30/11/-1 - 30/11/-1

The assembly of Vaccinia virus (VACV) takes place in the host cytoplasm and involves a unique mechanism of membrane biogenesis via the recruitment and remodelling of cellular membranes. Vesicles derived from the endoplasmic reticulum are ruptured open into membrane intermediates with stabilized open ends, which then associate into typical open VACV-crescents. Following packaging of viral core proteins and genome into the growing crescents, the immature virions are formed after closure but need to undergo further maturation steps to form infectious particles. Such a dynamic remodelling of cellular membranes is uncommon and involves not only fission and fusion but also the formation of vesicles derived from the endoplasmic reticulum and the maintenance of hydrophobic open ends in the host cytoplasm. Five viral membrane assembly proteins or VMAPs are known to be essential for the assembly of the viral membrane of VACV. In order to characterize the role of these proteins in remodelling membranes and how they interact with each other or additional partners, we would like to establish a cell-free expression system in combination with membrane-mimicking liposomes or microsomes in the lab.

- Cloning
- Protein expression (cell-free system)
- Protein purification (affinity purification, size-exclusion chromatography)
- Work with lipids (small unilamellar vesicles and microsomes)
- Electron microscopy (negative staining and cryo)

https://doi.org/10.1016/j.virol.2015.02.003
https://doi.org/10.1073/pnas.1716255114
http://dx.doi.org/10.1016/j.febslet.2014.05.061
https://doi.org/10.1038/nprot.2009.207

• Medicine and health sciences, Veterinary science
• Pharmacy, Biology, Physics and Chemistry