Unraveling the molecular mechanisms of Hantavirus assembly
Final Report Abstract
Hantaviruses (HV) can cause life-threatening human zoonoses with fatality rates up to 50%. Periodic HV outbreaks also occur in Germany, where up to 3000 cases are reported yearly by the Robert Koch Institute. However, the treatment of diseases caused by HV is currently only symptomatic and, so far, no targeted approach is available. The reason for this is mainly the lack of information about the molecular processes involved in the infection. The detailed replication mechanism of HV needs to be quantitatively characterized and understood to prevent future pandemics. Our work aimed at elucidating specific interactions between viral components (i.e., protein-protein interactions) that enable the assembly of new virions in infected cells. In particular, we have quantitatively investigated the interactions between the HV glycoproteins Gn and Gc in the context of the assembly process in living cells. To this aim, we have employed advanced fluorescence microscopy techniques (fluorescence fluctuation analysis) that can monitor multimerization and complex formation as a function of protein concentration and intracellular distribution. Our results show that Gn and Gc form up to tetramers and dimers, respectively. As a result of hetero-interactions, these glycoproteins form large multimers in the Golgi apparatus. Our experiments provided therefore a first experimental direct visualization of HV glycoprotein large-scale assembly in living cells. Furthermore, we have characterized the assembly behavior of the HV nucleoprotein NP and its interactions with several host factors (e.g., cytoskeleton) and the viral glycoproteins. Specifically, we were able to show that NP interacts with Gc and this interaction has a direct effect on the oligomeric state of Gc. In summary, our results contribute to the understanding of the molecular mechanisms of HV assembly and pave the road for further characterization of the interactions between viral proteins and host factors.
Publications
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Fluorescence microscopy methods for the study of protein oligomerization. Progress in Molecular Biology and Translational Science (2020), 1-41. American Geophysical Union (AGU).
Petazzi, Roberto Arturo; Aji, Amit Koikkarah & Chiantia, Salvatore
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Detection of Envelope Glycoprotein Assembly from Old World Hantaviruses in the Golgi Apparatus of Living Cells. Journal of Virology, 95(4).
Petazzi, R. A.; Koikkarah, A. A.; Tischler, N. D. & Chiantia, S.
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Characterization of Hantavirus N Protein Intracellular Dynamics and Localization. Viruses, 14(3), 457.
Welke, Robert-William; Sperber, Hannah Sabeth; Bergmann, Ronny; Koikkarah, Amit; Menke, Laura; Sieben, Christian; Krüger, Detlev H.; Chiantia, Salvatore; Herrmann, Andreas & Schwarzer, Roland
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Advances in fluorescence microscopy for orthohantavirus research. Microscopy, 72(3), 191-203.
Menke, Laura; Sperber, Hannah S; Aji, Amit Koikkarah; Chiantia, Salvatore; Schwarzer, Roland & Sieben, Christian
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Benchmarking of novel green fluorescent proteins for the quantification of protein oligomerization in living cells. (2023, 2, 7). American Geophysical Union (AGU).
Petrich, Annett; Aji, Amit Koikkarah; Dunsing, Valentin & Chiantia, Salvatore
