| Popis |
Human echovirus 18 is a member of the Enterovirus B species. It has diverse tissue tropism: the cells of the nervous system, lower respiratory tract, and gastrointestinal system. The long-term effects of viral encephalitis are acquired brain injury, memory loss, and altered consciousness [1]. Furthermore, it has been reported that this virus is connected to chronic gastrointestinal diseases, particularly, to Crohn’s disease [2]. In this project, we characterize the interactions of cell and Echovirus 18 particles and the genome delivery in situ. We study the replication cycle to determine the mechanisms of recombination that cause the emergence of new viral strains. By observation of the virion assembly, we identify the mechanism of genome packaging. We use the focused-ion beam milling (FIBM) to prepare lamellae of the infected cells and image them using cryo-electron microscopy. The thin (200 to 300 nm) lamellae are used to record tomographic tilt series [3, 4]. The image alignment is followed by the 3D reconstruction of cell sections [5]. The reconstructed parts of cells provide information about the positioning and interactions of the macromolecular complexes of the cell with the virion assemblies and their intermediate states. Since enteroviruses are a homogeneous group of viruses, the data for human enterovirus 18 will be relevant also as a reference for further experiments on other enterovirus species. 1. M. C. Huang, S. M. Wang, Y. W. Hsu, H. C. Lin, C. Y. Chi, C. C. Liu, Pediatrics, 118, (2006), e1785. 2. J. Chia, A. Chia, D. Wang, R. El-Habbal, S. Jyotheeswaran, E. McCloud, Open J Gastroenterol, 9, (2019) 174. 3. A. Rigort, F. J. Bäuerlein, E. Villa, M. Eibauer, T. Laugks, W. Baumeister, J. M. Plitzko, Proc Natl Acad Sci U.S.A., 109, (2012), 4449. 4. M. Schaffer, J. Mahamid, B. D. Engel, T. Laugks, W. Baumeister, J. M. Plitzko, J Struct Biol, 197 (2017), 73. 5. J. M. Plitzko, B. Schuler, P. Selenko, Curr Opin Struc Biol, 46, (2017), 110. We acknowledge the Cryo-electron microscopy and Tomography Core Facility of CEITEC MU for their support in data collection and analysis.
|