Structure of tick-borne encephalitis virus immature particle solved by cryo-electron microscopy and sub-tomogram averaging

Tibor Füzik1, Lenka Šmerdová1, Petra Pokorná Formanová2, Petra Straková2, Jiří Nováček3, Daniel Růžek2, Pavel Plevka1

1 Structural Biology, Central European Institute of Technology, Masaryk University, Kamenice 5, Brno, Czech Republic, tibor.fuzik@ceitec.muni.cz

2Department of Virology, Veterinary Research Institute, Hudcova 70, Brno, Czech Republic

3Cryo-electron Microscopy and Tomography Core Facility, Central European Institute of Technology, Masaryk University, Kamenice 5, Brno, Czech Republic

Tick-borne encephalitis virus (TBEV) is an enveloped virus belonging to the family Flaviviridae. It is mainly transmitted by ticks and causes severe disease of central nervous system in humans. Virion surface is covered by envelope proteins (E-protein), that are together with the membrane proteins (M-protein) anchored in virus lipid bilayer. During the viral life cycle, the immature non-infectious virus undergoes a maturation process. This process includes proteolytic cleavage of prM and major reorganization of the envelope proteins on the viral surface.

To determine the structure of immature TBEV particles, we purified them from infected tissue culture cells and used cryo-electron microscopy for visualization. In comparison with smooth mature TBEV particles, the immature particles have “spiky” surface formed by the E-protein-prM-protein complex. We combined cryo-electron tomography and sub-tomogram averaging with single-particle analysis methods using localized reconstruction of the surface “spikes”, to determine the high-resolution structure of the immature E-protein complexes and their interaction with the prM-protein. The organization of the particle surface indicates that the TBEV maturation mechanism involves a complex reorganization of the envelope proteins on the viral surface.

The results show more detailed insight in the viral maturation process which may be targeted by specific antiviral drugs.