Characterization of Hantavirus N Protein Intracellular Dynamics and Localization
Robert-William Welke,
Hannah Sabeth Sperber,
Ronny Bergmann,
Amit Koikkarah,
Laura Menke,
Christian Sieben,
Detlev H. Krüger,
Salvatore Chiantia,
Andreas Herrmann,
Roland Schwarzer
Affiliations
Robert-William Welke
Department of Molecular Biophysics, Humboldt University, 10115 Berlin, Germany
Hannah Sabeth Sperber
Institute for Translational HIV Research, University Hospital Essen, 45147 Essen, Germany
Ronny Bergmann
Department of Molecular Biophysics, Humboldt University, 10115 Berlin, Germany
Amit Koikkarah
Institute of Biochemistry and Biology, University of Potsdam, 14476 Potsdam, Germany
Laura Menke
Nanoscale Infection Biology Group, Department of Cell Biology, Helmholtz Centre for Infection Research, 38124 Braunschweig, Germany
Christian Sieben
Nanoscale Infection Biology Group, Department of Cell Biology, Helmholtz Centre for Infection Research, 38124 Braunschweig, Germany
Detlev H. Krüger
Institut für Virologie, Charité–Universitätsmedizin Berlin, Gliedkörperschaft der Freien Universität Berlin und der Humboldt-Universität zu Berlin, 10117 Berlin, Germany
Salvatore Chiantia
Institute of Biochemistry and Biology, University of Potsdam, 14476 Potsdam, Germany
Andreas Herrmann
Department of Molecular Biophysics, Humboldt University, 10115 Berlin, Germany
Roland Schwarzer
Institute for Translational HIV Research, University Hospital Essen, 45147 Essen, Germany
Hantaviruses are enveloped viruses that possess a tri-segmented, negative-sense RNA genome. The viral S-segment encodes the multifunctional nucleocapsid protein (N), which is involved in genome packaging, intracellular protein transport, immunoregulation, and several other crucial processes during hantavirus infection. In this study, we generated fluorescently tagged N protein constructs derived from Puumalavirus (PUUV), the dominant hantavirus species in Central, Northern, and Eastern Europe. We comprehensively characterized this protein in the rodent cell line CHO-K1, monitoring the dynamics of N protein complex formation and investigating co-localization with host proteins as well as the viral glycoproteins Gc and Gn. We observed formation of large, fibrillar PUUV N protein aggregates, rapidly coalescing from early punctate and spike-like assemblies. Moreover, we found significant spatial correlation of N with vimentin, actin, and P-bodies but not with microtubules. N constructs also co-localized with Gn and Gc albeit not as strongly as the glycoproteins associated with each other. Finally, we assessed oligomerization of N constructs, observing efficient and concentration-dependent multimerization, with complexes comprising more than 10 individual proteins.
