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

Candida da Fonseca Pereira, Paula Clarisa Ellenberg, Kate L Jones, Tara L Fernandez, Redmond P Smyth, David J Hawkes, Marcel Hijnen, Valerie Vivet-Boudou, Roland Marquet, Iain Johnson, Johnson Mak

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Abstract

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.
Original languageEnglish
Article numbere17016
Pages (from-to)1 - 10
Number of pages10
JournalPLoS ONE
Volume6
Issue number2
DOIs
Publication statusPublished - 2011

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