Biochemistry; Cell Membrane Structures; Cryoelectron Microscopy; Electrophoresis, Capillary; Endosomes; Membrane Lipids; Molecular Biology; Rhinovirus; RNA, Viral; Spectrometry, Fluorescence
Working with human rhinoviruses (HRVs) that lack a lipid membrane and are the predominant cause of common colds, we aim at identifying so far unknown viral receptors, the different mechanisms underlying viral uptake, the process of genome release and the structural basis of the transfer of the viral genome through lipid membranes. For some viruses there is experimental support for RNA transfer between endosomal and cytoplasmic compartments which, however, has so far not been demonstrated explicitly and the putative membrane pore has not been visualized.
We developed a liposomal in vitro system mimicking the transfer of the viral RNA through the endosomal membrane that is currently being used for structural analysis. Within the frame of several international collaborations, solving the 3D structure of subviral particles is underway.
Techniques, methods & infrastructure
We address these questions by using biochemical, molecular biological, biophysical, and structural biology techniques, such as selection and characterization of viral mutants, expression library screening, fluorescence correlation spectroscopy, capillary electrophoresis, and cryo-electron microscopy.
- Ganjian, H. et al., 2017. ICAM-1 Binding Rhinoviruses Enter HeLa Cells via Multiple Pathways and Travel to Distinct Intracellular Compartments for Uncoating. Viruses, 9(4), p.68. Available at: http://dx.doi.org/10.3390/v9040068.
- Otahal, A. et al., 2015. Release of Vesicular Stomatitis Virus Spike Protein G-Pseudotyped Lentivirus from the Host Cell Is Impaired upon Low-Density Lipoprotein Receptor Overexpression D. S. Lyles, ed. Journal of Virology, 89(22), pp.11723-11726. Available at: http://dx.doi.org/10.1128/JVI.01869-15.
- Harutyunyan, S., Kowalski, H. & Blaas, D., 2014. The Rhinovirus Subviral A-Particle Exposes 3�-Terminal Sequences of Its Genomic RNA. Journal of Virology, 88(11), pp.6307-6317. Available at: http://dx.doi.org/10.1128/JVI.00539-14.
- Pickl-Herk, A. et al., 2013. Uncoating of common cold virus is preceded by RNA switching as determined by X-ray and cryo-EM analyses of the subviral A-particle. Proceedings of the National Academy of Sciences, 110(50), pp.20063-20068. Available at: http://dx.doi.org/10.1073/pnas.1312128110.
- Harutyunyan, S. et al., 2013. Viral Uncoating Is Directional: Exit of the Genomic RNA in a Common Cold Virus Starts with the Poly-(A) Tail at the 3�-End F. A. Rey, ed. PLoS Pathogens, 9(4), p.e1003270. Available at: http://dx.doi.org/10.1371/journal.ppat.1003270.