RNAi Screening Reveals Proteasome- and Cullin3-Dependent Stages in Vaccinia Virus Infection
A two-step, automated, high-throughput RNAi silencing screen was used to identify host cell factors required during vaccinia virus infection. Validation and analysis of clustered hits revealed previously unknown processes during virus entry, including a mechanism for genome uncoating. Viral core pro...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2012-10-01
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| Series: | Cell Reports |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2211124712002847 |
| _version_ | 1811315095646502912 |
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| author | Jason Mercer Berend Snijder Raphael Sacher Christine Burkard Christopher Karl Ernst Bleck Henning Stahlberg Lucas Pelkmans Ari Helenius |
| author_facet | Jason Mercer Berend Snijder Raphael Sacher Christine Burkard Christopher Karl Ernst Bleck Henning Stahlberg Lucas Pelkmans Ari Helenius |
| author_sort | Jason Mercer |
| collection | DOAJ |
| description | A two-step, automated, high-throughput RNAi silencing screen was used to identify host cell factors required during vaccinia virus infection. Validation and analysis of clustered hits revealed previously unknown processes during virus entry, including a mechanism for genome uncoating. Viral core proteins were found to be already ubiquitinated during virus assembly. After entering the cytosol of an uninfected cell, the viral DNA was released from the core through the activity of the cell’s proteasomes. Next, a Cullin3-based ubiquitin ligase mediated a further round of ubiquitination and proteasome action. This was needed in order to initiate viral DNA replication. The results accentuate the value of large-scale RNAi screens in providing directions for detailed cell biological investigation of complex pathways. The list of cell functions required during poxvirus infection will, moreover, provide a resource for future virus-host cell interaction studies and for the discovery of antivirals. |
| first_indexed | 2024-04-13T11:24:10Z |
| format | Article |
| id | doaj.art-47bd2d1f31304c188aae57965f785516 |
| institution | Directory Open Access Journal |
| issn | 2211-1247 |
| language | English |
| last_indexed | 2024-04-13T11:24:10Z |
| publishDate | 2012-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Cell Reports |
| spelling | doaj.art-47bd2d1f31304c188aae57965f7855162022-12-22T02:48:45ZengElsevierCell Reports2211-12472012-10-01241036104710.1016/j.celrep.2012.09.003RNAi Screening Reveals Proteasome- and Cullin3-Dependent Stages in Vaccinia Virus InfectionJason Mercer0Berend Snijder1Raphael Sacher2Christine Burkard3Christopher Karl Ernst Bleck4Henning Stahlberg5Lucas Pelkmans6Ari Helenius7Institute of Biochemistry, ETH Zürich, 8093 Zürich, SwitzerlandInstitute of Molecular Life Sciences, University of Zürich, 8057 Zürich, SwitzerlandInstitute of Molecular Systems Biology, ETH Zürich, 8093 Zürich, SwitzerlandInstitute of Biochemistry, ETH Zürich, 8093 Zürich, SwitzerlandCenter for Cellular Imaging and NanoAnalytics (C-CINA), Structural Biology and Biophysics, University Basel, Biozentrum, 4058 Basel, SwitzerlandCenter for Cellular Imaging and NanoAnalytics (C-CINA), Structural Biology and Biophysics, University Basel, Biozentrum, 4058 Basel, SwitzerlandInstitute of Molecular Life Sciences, University of Zürich, 8057 Zürich, SwitzerlandInstitute of Biochemistry, ETH Zürich, 8093 Zürich, SwitzerlandA two-step, automated, high-throughput RNAi silencing screen was used to identify host cell factors required during vaccinia virus infection. Validation and analysis of clustered hits revealed previously unknown processes during virus entry, including a mechanism for genome uncoating. Viral core proteins were found to be already ubiquitinated during virus assembly. After entering the cytosol of an uninfected cell, the viral DNA was released from the core through the activity of the cell’s proteasomes. Next, a Cullin3-based ubiquitin ligase mediated a further round of ubiquitination and proteasome action. This was needed in order to initiate viral DNA replication. The results accentuate the value of large-scale RNAi screens in providing directions for detailed cell biological investigation of complex pathways. The list of cell functions required during poxvirus infection will, moreover, provide a resource for future virus-host cell interaction studies and for the discovery of antivirals.http://www.sciencedirect.com/science/article/pii/S2211124712002847 |
| spellingShingle | Jason Mercer Berend Snijder Raphael Sacher Christine Burkard Christopher Karl Ernst Bleck Henning Stahlberg Lucas Pelkmans Ari Helenius RNAi Screening Reveals Proteasome- and Cullin3-Dependent Stages in Vaccinia Virus Infection Cell Reports |
| title | RNAi Screening Reveals Proteasome- and Cullin3-Dependent Stages in Vaccinia Virus Infection |
| title_full | RNAi Screening Reveals Proteasome- and Cullin3-Dependent Stages in Vaccinia Virus Infection |
| title_fullStr | RNAi Screening Reveals Proteasome- and Cullin3-Dependent Stages in Vaccinia Virus Infection |
| title_full_unstemmed | RNAi Screening Reveals Proteasome- and Cullin3-Dependent Stages in Vaccinia Virus Infection |
| title_short | RNAi Screening Reveals Proteasome- and Cullin3-Dependent Stages in Vaccinia Virus Infection |
| title_sort | rnai screening reveals proteasome and cullin3 dependent stages in vaccinia virus infection |
| url | http://www.sciencedirect.com/science/article/pii/S2211124712002847 |
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