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Labeling of multiple HIV-1 proteins with the biarsenical-tetracysteine system

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journal contribution
posted on 2011-01-01, 00:00 authored by C Pereira, P Ellenberg, K Jones, T Fernandez, R Smyth, D Hawkes, M Hijnen, V Vivet-Boudou, R Marquet, I Johnson, Johnson Mak
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.

History

Journal

PLoS one

Volume

6

Pagination

1 - 10

Location

San Francisco, Calif.

Open access

  • Yes

ISSN

1932-6203

Language

eng

Publication classification

C1.1 Refereed article in a scholarly journal

Copyright notice

2011, Public Library of Science