Dissecting virus entry: replication-independent analysis of virus binding, internalization, and penetration using minimal complementation of β-galactosidase.
Studies of viral entry into host cells often rely on the detection of post-entry parameters, such as viral replication or the expression of a reporter gene, rather than on measuring entry per se. The lack of assays to easily detect the different steps of entry severely hampers the analysis of this k...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2014-01-01
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| Series: | PLoS ONE |
| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/25025332/pdf/?tool=EBI |
| _version_ | 1819181027011067904 |
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| author | Christine Burkard Louis-Marie Bloyet Oliver Wicht Frank J van Kuppeveld Peter J M Rottier Cornelis A M de Haan Berend Jan Bosch |
| author_facet | Christine Burkard Louis-Marie Bloyet Oliver Wicht Frank J van Kuppeveld Peter J M Rottier Cornelis A M de Haan Berend Jan Bosch |
| author_sort | Christine Burkard |
| collection | DOAJ |
| description | Studies of viral entry into host cells often rely on the detection of post-entry parameters, such as viral replication or the expression of a reporter gene, rather than on measuring entry per se. The lack of assays to easily detect the different steps of entry severely hampers the analysis of this key process in virus infection. Here we describe novel, highly adaptable viral entry assays making use of minimal complementation of the E. coli β-galactosidase in mammalian cells. Enzyme activity is reconstituted when a small intravirion peptide (α-peptide) is complementing the inactive mutant form ΔM15 of β-galactosidase. The method allows to dissect and to independently detect binding, internalization, and fusion of viruses during host cell entry. Here we use it to confirm and extend current knowledge on the entry process of two enveloped viruses: vesicular stomatitis virus (VSV) and murine hepatitis coronavirus (MHV). |
| first_indexed | 2024-12-22T22:23:42Z |
| format | Article |
| id | doaj.art-39de291686d94fdaabb6cf7a2379338a |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-22T22:23:42Z |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-39de291686d94fdaabb6cf7a2379338a2022-12-21T18:10:36ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0197e10176210.1371/journal.pone.0101762Dissecting virus entry: replication-independent analysis of virus binding, internalization, and penetration using minimal complementation of β-galactosidase.Christine BurkardLouis-Marie BloyetOliver WichtFrank J van KuppeveldPeter J M RottierCornelis A M de HaanBerend Jan BoschStudies of viral entry into host cells often rely on the detection of post-entry parameters, such as viral replication or the expression of a reporter gene, rather than on measuring entry per se. The lack of assays to easily detect the different steps of entry severely hampers the analysis of this key process in virus infection. Here we describe novel, highly adaptable viral entry assays making use of minimal complementation of the E. coli β-galactosidase in mammalian cells. Enzyme activity is reconstituted when a small intravirion peptide (α-peptide) is complementing the inactive mutant form ΔM15 of β-galactosidase. The method allows to dissect and to independently detect binding, internalization, and fusion of viruses during host cell entry. Here we use it to confirm and extend current knowledge on the entry process of two enveloped viruses: vesicular stomatitis virus (VSV) and murine hepatitis coronavirus (MHV).https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/25025332/pdf/?tool=EBI |
| spellingShingle | Christine Burkard Louis-Marie Bloyet Oliver Wicht Frank J van Kuppeveld Peter J M Rottier Cornelis A M de Haan Berend Jan Bosch Dissecting virus entry: replication-independent analysis of virus binding, internalization, and penetration using minimal complementation of β-galactosidase. PLoS ONE |
| title | Dissecting virus entry: replication-independent analysis of virus binding, internalization, and penetration using minimal complementation of β-galactosidase. |
| title_full | Dissecting virus entry: replication-independent analysis of virus binding, internalization, and penetration using minimal complementation of β-galactosidase. |
| title_fullStr | Dissecting virus entry: replication-independent analysis of virus binding, internalization, and penetration using minimal complementation of β-galactosidase. |
| title_full_unstemmed | Dissecting virus entry: replication-independent analysis of virus binding, internalization, and penetration using minimal complementation of β-galactosidase. |
| title_short | Dissecting virus entry: replication-independent analysis of virus binding, internalization, and penetration using minimal complementation of β-galactosidase. |
| title_sort | dissecting virus entry replication independent analysis of virus binding internalization and penetration using minimal complementation of β galactosidase |
| url | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/25025332/pdf/?tool=EBI |
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