Nuclear envelope impairment is facilitated by the herpes simplex virus 1 Us3 kinase [version 1; peer review: 2 approved]
Background: Capsids of herpes simplex virus 1 (HSV-1) are assembled in the nucleus, translocated either to the perinuclear space by budding at the inner nuclear membrane acquiring tegument and envelope, or released to the cytosol in a “naked” state via impaired nuclear pores that finally results in...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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F1000 Research Ltd
2019-02-01
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| Series: | F1000Research |
| Online Access: | https://f1000research.com/articles/8-198/v1 |
| _version_ | 1828824979607650304 |
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| author | Peter Wild Sabine Leisinger Anna Paula de Oliveira Jana Doehner Elisabeth M. Schraner Cornel Fraevel Mathias Ackermann Andres Kaech |
| author_facet | Peter Wild Sabine Leisinger Anna Paula de Oliveira Jana Doehner Elisabeth M. Schraner Cornel Fraevel Mathias Ackermann Andres Kaech |
| author_sort | Peter Wild |
| collection | DOAJ |
| description | Background: Capsids of herpes simplex virus 1 (HSV-1) are assembled in the nucleus, translocated either to the perinuclear space by budding at the inner nuclear membrane acquiring tegument and envelope, or released to the cytosol in a “naked” state via impaired nuclear pores that finally results in impairment of the nuclear envelope. The Us3 gene encodes a protein acting as a kinase, which is responsible for phosphorylation of numerous viral and cellular substrates. The Us3 kinase plays a crucial role in nucleus to cytoplasm capsid translocation. We thus investigate the nuclear surface in order to evaluate the significance of Us3 in maintenance of the nuclear envelope during HSV-1 infection. Methods: To address alterations of the nuclear envelope and capsid nucleus to cytoplasm translocation related to the function of the Us3 kinase we investigated cells infected with wild type HSV-1 or the Us3 deletion mutant R7041(∆Us3) by transmission electron microscopy, focused ion-beam electron scanning microscopy, cryo-field emission scanning electron microscopy, confocal super resolution light microscopy, and polyacrylamide gel electrophoresis. Results: Confocal super resolution microscopy and cryo-field emission scanning electron microscopy revealed decrement in pore numbers in infected cells. Number and degree of pore impairment was significantly reduced after infection with R7041(∆Us3) compared to infection with wild type HSV-1. The nuclear surface was significantly enlarged in cells infected with any of the viruses. Morphometric analysis revealed that additional nuclear membranes were produced forming multiple folds and caveolae, in which virions accumulated as documented by three-dimensional reconstruction after ion-beam scanning electron microscopy. Finally, significantly more R7041(∆Us3) capsids were retained in the nucleus than wild-type capsids whereas the number of R7041(∆Us3) capsids in the cytosol was significantly lower. Conclusions: The data indicate that Us3 kinase is involved in facilitation of nuclear pore impairment and, concomitantly, in capsid release through impaired nuclear envelope. |
| first_indexed | 2024-12-12T14:07:22Z |
| format | Article |
| id | doaj.art-a89a31c4502e4614a812542be8c74f34 |
| institution | Directory Open Access Journal |
| issn | 2046-1402 |
| language | English |
| last_indexed | 2024-12-12T14:07:22Z |
| publishDate | 2019-02-01 |
| publisher | F1000 Research Ltd |
| record_format | Article |
| series | F1000Research |
| spelling | doaj.art-a89a31c4502e4614a812542be8c74f342022-12-22T00:22:10ZengF1000 Research LtdF1000Research2046-14022019-02-01810.12688/f1000research.17802.119463Nuclear envelope impairment is facilitated by the herpes simplex virus 1 Us3 kinase [version 1; peer review: 2 approved]Peter Wild0Sabine Leisinger1Anna Paula de Oliveira2Jana Doehner3Elisabeth M. Schraner4Cornel Fraevel5Mathias Ackermann6Andres Kaech7Department of Veterinary Anatomy, University of Zuerich, Zürich, CH-8057, SwitzerlandDepartment of Veterinary Anatomy, University of Zuerich, Zürich, CH-8057, SwitzerlandInstute of Virology, University of Zürich, Zürich, ZH-8057, SwitzerlandCenter for Microcopy and Image Analysis, Universit of Zürich, Zürich, CH-8057, SwitzerlandInstute of Virology, University of Zürich, Zürich, ZH-8057, SwitzerlandInstute of Virology, University of Zürich, Zürich, ZH-8057, SwitzerlandInstute of Virology, University of Zürich, Zürich, ZH-8057, SwitzerlandCenter for Microcopy and Image Analysis, Universit of Zürich, Zürich, CH-8057, SwitzerlandBackground: Capsids of herpes simplex virus 1 (HSV-1) are assembled in the nucleus, translocated either to the perinuclear space by budding at the inner nuclear membrane acquiring tegument and envelope, or released to the cytosol in a “naked” state via impaired nuclear pores that finally results in impairment of the nuclear envelope. The Us3 gene encodes a protein acting as a kinase, which is responsible for phosphorylation of numerous viral and cellular substrates. The Us3 kinase plays a crucial role in nucleus to cytoplasm capsid translocation. We thus investigate the nuclear surface in order to evaluate the significance of Us3 in maintenance of the nuclear envelope during HSV-1 infection. Methods: To address alterations of the nuclear envelope and capsid nucleus to cytoplasm translocation related to the function of the Us3 kinase we investigated cells infected with wild type HSV-1 or the Us3 deletion mutant R7041(∆Us3) by transmission electron microscopy, focused ion-beam electron scanning microscopy, cryo-field emission scanning electron microscopy, confocal super resolution light microscopy, and polyacrylamide gel electrophoresis. Results: Confocal super resolution microscopy and cryo-field emission scanning electron microscopy revealed decrement in pore numbers in infected cells. Number and degree of pore impairment was significantly reduced after infection with R7041(∆Us3) compared to infection with wild type HSV-1. The nuclear surface was significantly enlarged in cells infected with any of the viruses. Morphometric analysis revealed that additional nuclear membranes were produced forming multiple folds and caveolae, in which virions accumulated as documented by three-dimensional reconstruction after ion-beam scanning electron microscopy. Finally, significantly more R7041(∆Us3) capsids were retained in the nucleus than wild-type capsids whereas the number of R7041(∆Us3) capsids in the cytosol was significantly lower. Conclusions: The data indicate that Us3 kinase is involved in facilitation of nuclear pore impairment and, concomitantly, in capsid release through impaired nuclear envelope.https://f1000research.com/articles/8-198/v1 |
| spellingShingle | Peter Wild Sabine Leisinger Anna Paula de Oliveira Jana Doehner Elisabeth M. Schraner Cornel Fraevel Mathias Ackermann Andres Kaech Nuclear envelope impairment is facilitated by the herpes simplex virus 1 Us3 kinase [version 1; peer review: 2 approved] F1000Research |
| title | Nuclear envelope impairment is facilitated by the herpes simplex virus 1 Us3 kinase [version 1; peer review: 2 approved] |
| title_full | Nuclear envelope impairment is facilitated by the herpes simplex virus 1 Us3 kinase [version 1; peer review: 2 approved] |
| title_fullStr | Nuclear envelope impairment is facilitated by the herpes simplex virus 1 Us3 kinase [version 1; peer review: 2 approved] |
| title_full_unstemmed | Nuclear envelope impairment is facilitated by the herpes simplex virus 1 Us3 kinase [version 1; peer review: 2 approved] |
| title_short | Nuclear envelope impairment is facilitated by the herpes simplex virus 1 Us3 kinase [version 1; peer review: 2 approved] |
| title_sort | nuclear envelope impairment is facilitated by the herpes simplex virus 1 us3 kinase version 1 peer review 2 approved |
| url | https://f1000research.com/articles/8-198/v1 |
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