Characterization of Hantavirus N Protein Intracellular Dynamics and Localization
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 h...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-02-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/14/3/457 |
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| author | Robert-William Welke Hannah Sabeth Sperber Ronny Bergmann Amit Koikkarah Laura Menke Christian Sieben Detlev H. Krüger Salvatore Chiantia Andreas Herrmann Roland Schwarzer |
| author_facet | Robert-William Welke Hannah Sabeth Sperber Ronny Bergmann Amit Koikkarah Laura Menke Christian Sieben Detlev H. Krüger Salvatore Chiantia Andreas Herrmann Roland Schwarzer |
| author_sort | Robert-William Welke |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-09T12:19:20Z |
| format | Article |
| id | doaj.art-e3d399b345e347d190ea0b523cd2c36a |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T12:19:20Z |
| publishDate | 2022-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-e3d399b345e347d190ea0b523cd2c36a2023-11-30T22:43:44ZengMDPI AGViruses1999-49152022-02-0114345710.3390/v14030457Characterization of Hantavirus N Protein Intracellular Dynamics and LocalizationRobert-William Welke0Hannah Sabeth Sperber1Ronny Bergmann2Amit Koikkarah3Laura Menke4Christian Sieben5Detlev H. Krüger6Salvatore Chiantia7Andreas Herrmann8Roland Schwarzer9Department of Molecular Biophysics, Humboldt University, 10115 Berlin, GermanyInstitute for Translational HIV Research, University Hospital Essen, 45147 Essen, GermanyDepartment of Molecular Biophysics, Humboldt University, 10115 Berlin, GermanyInstitute of Biochemistry and Biology, University of Potsdam, 14476 Potsdam, GermanyNanoscale Infection Biology Group, Department of Cell Biology, Helmholtz Centre for Infection Research, 38124 Braunschweig, GermanyNanoscale Infection Biology Group, Department of Cell Biology, Helmholtz Centre for Infection Research, 38124 Braunschweig, GermanyInstitut für Virologie, Charité–Universitätsmedizin Berlin, Gliedkörperschaft der Freien Universität Berlin und der Humboldt-Universität zu Berlin, 10117 Berlin, GermanyInstitute of Biochemistry and Biology, University of Potsdam, 14476 Potsdam, GermanyDepartment of Molecular Biophysics, Humboldt University, 10115 Berlin, GermanyInstitute for Translational HIV Research, University Hospital Essen, 45147 Essen, GermanyHantaviruses 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.https://www.mdpi.com/1999-4915/14/3/457hantavirusN proteinoligomerizationactinP-bodiesvimentin |
| spellingShingle | Robert-William Welke Hannah Sabeth Sperber Ronny Bergmann Amit Koikkarah Laura Menke Christian Sieben Detlev H. Krüger Salvatore Chiantia Andreas Herrmann Roland Schwarzer Characterization of Hantavirus N Protein Intracellular Dynamics and Localization Viruses hantavirus N protein oligomerization actin P-bodies vimentin |
| title | Characterization of Hantavirus N Protein Intracellular Dynamics and Localization |
| title_full | Characterization of Hantavirus N Protein Intracellular Dynamics and Localization |
| title_fullStr | Characterization of Hantavirus N Protein Intracellular Dynamics and Localization |
| title_full_unstemmed | Characterization of Hantavirus N Protein Intracellular Dynamics and Localization |
| title_short | Characterization of Hantavirus N Protein Intracellular Dynamics and Localization |
| title_sort | characterization of hantavirus n protein intracellular dynamics and localization |
| topic | hantavirus N protein oligomerization actin P-bodies vimentin |
| url | https://www.mdpi.com/1999-4915/14/3/457 |
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