Labeling of multiple HIV-1 proteins with the biarsenical-tetracysteine system.
Due to its small size and versatility, the biarsenical-tetracysteine system is an attractive way to label viral proteins for live cell imaging. This study describes the genetic labeling of the human immunodeficiency virus type 1 (HIV-1) structural proteins (matrix, capsid and nucleocapsid), enzymes...
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2011
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oai:doaj.org-article:0c04dcbf311646b9973335f0827940222021-11-18T06:58:53ZLabeling of multiple HIV-1 proteins with the biarsenical-tetracysteine system.1932-620310.1371/journal.pone.0017016https://doaj.org/article/0c04dcbf311646b9973335f0827940222011-02-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21347302/?tool=EBIhttps://doaj.org/toc/1932-6203Due to its small size and versatility, the biarsenical-tetracysteine system is an attractive way to label viral proteins for live cell imaging. This study describes the genetic labeling of the human immunodeficiency virus type 1 (HIV-1) structural proteins (matrix, capsid and nucleocapsid), enzymes (protease, reverse transcriptase, RNAse H and integrase) and envelope glycoprotein 120 with a tetracysteine tag in the context of a full-length virus. We measure the impact of these modifications on the natural virus infection and, most importantly, present the first infectious HIV-1 construct containing a fluorescently-labeled nucleocapsid protein. Furthermore, due to the high background levels normally associated with the labeling of tetracysteine-tagged proteins we have also optimized a metabolic labeling system that produces infectious virus containing the natural envelope glycoproteins and specifically labeled tetracysteine-tagged proteins that can easily be detected after virus infection of T-lymphocytes. This approach can be adapted to other viral systems for the visualization of the interplay between virus and host cell during infection.Cândida F PereiraPaula C EllenbergKate L JonesTara L FernandezRedmond P SmythDavid J HawkesMarcel HijnenValérie Vivet-BoudouRoland MarquetIain JohnsonJohnson MakPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 2, p e17016 (2011) |
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| collection |
DOAJ |
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EN |
| topic |
Medicine R Science Q |
| spellingShingle |
Medicine R Science Q Cândida F Pereira Paula C Ellenberg Kate L Jones Tara L Fernandez Redmond P Smyth David J Hawkes Marcel Hijnen Valérie Vivet-Boudou Roland Marquet Iain Johnson Johnson Mak Labeling of multiple HIV-1 proteins with the biarsenical-tetracysteine system. |
| description |
Due to its small size and versatility, the biarsenical-tetracysteine system is an attractive way to label viral proteins for live cell imaging. This study describes the genetic labeling of the human immunodeficiency virus type 1 (HIV-1) structural proteins (matrix, capsid and nucleocapsid), enzymes (protease, reverse transcriptase, RNAse H and integrase) and envelope glycoprotein 120 with a tetracysteine tag in the context of a full-length virus. We measure the impact of these modifications on the natural virus infection and, most importantly, present the first infectious HIV-1 construct containing a fluorescently-labeled nucleocapsid protein. Furthermore, due to the high background levels normally associated with the labeling of tetracysteine-tagged proteins we have also optimized a metabolic labeling system that produces infectious virus containing the natural envelope glycoproteins and specifically labeled tetracysteine-tagged proteins that can easily be detected after virus infection of T-lymphocytes. This approach can be adapted to other viral systems for the visualization of the interplay between virus and host cell during infection. |
| format |
article |
| author |
Cândida F Pereira Paula C Ellenberg Kate L Jones Tara L Fernandez Redmond P Smyth David J Hawkes Marcel Hijnen Valérie Vivet-Boudou Roland Marquet Iain Johnson Johnson Mak |
| author_facet |
Cândida F Pereira Paula C Ellenberg Kate L Jones Tara L Fernandez Redmond P Smyth David J Hawkes Marcel Hijnen Valérie Vivet-Boudou Roland Marquet Iain Johnson Johnson Mak |
| author_sort |
Cândida F Pereira |
| title |
Labeling of multiple HIV-1 proteins with the biarsenical-tetracysteine system. |
| title_short |
Labeling of multiple HIV-1 proteins with the biarsenical-tetracysteine system. |
| title_full |
Labeling of multiple HIV-1 proteins with the biarsenical-tetracysteine system. |
| title_fullStr |
Labeling of multiple HIV-1 proteins with the biarsenical-tetracysteine system. |
| title_full_unstemmed |
Labeling of multiple HIV-1 proteins with the biarsenical-tetracysteine system. |
| title_sort |
labeling of multiple hiv-1 proteins with the biarsenical-tetracysteine system. |
| publisher |
Public Library of Science (PLoS) |
| publishDate |
2011 |
| url |
https://doaj.org/article/0c04dcbf311646b9973335f082794022 |
| work_keys_str_mv |
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