Proteomic analysis of HIV-1 Gag interacting partners using proximity-dependent biotinylation.

Valerie Le Sage,Andrew J Mouland,Fernando Valiente-Echeverría,Alessandro Cinti

doi:10.1186/s12985-015-0365-6

Valerie Le Sage, Andrew J Mouland + Show 2 more

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https://doi.org/10.1186/s12985-015-0365-6

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Abstract

BackgroundThe human immunodeficiency virus type 1 (HIV-1) Gag polyprotein is necessary and sufficient to assemble non-infectious particles. Given that HIV-1 subverts many host proteins at all stages of its life cycle, it is essential to identify these interactions as potential targets for antiretroviral therapy.FindingsThis work demonstrates the use of proximity-dependent biotin identification (BioID) of host proteins and complexes that are proximal to the N-terminal domains of the HIV-1 Gag polyprotein. Two of the hits identified in the BioID screen were validated by immunoprecipation and confirmed the interaction of DDX17 and RPS6 with HIV-1 Gag.ConclusionsOur results show that BioID is both a successful and complementary method to screen for nearby interacting proteins of HIV-1 Gag during the replicative cycle in different cell lines.Electronic supplementary materialThe online version of this article (doi:10.1186/s12985-015-0365-6) contains supplementary material, which is available to authorized users.

Highlights

The human immunodeficiency virus type 1 (HIV-1) Gag polyprotein is necessary and sufficient to assemble non-infectious particles
Our results show that biotin identification (BioID) is both a successful and complementary method to screen for nearby interacting proteins of HIV-1 Gag during the replicative cycle in different cell lines
The integrated proviral DNA is transcribed to generate multiply and singly spliced and full-length messenger RNAs. Viral RNA (vRNA) acts as both the mRNA for the Gag and Gag-Pol polyprotein precursors, and the genome that is encapsidated into assembling particles [1]

Summary

Introduction

The human immunodeficiency virus type 1 (HIV-1) Gag polyprotein is necessary and sufficient to assemble non-infectious particles. Conclusions: Our results show that BioID is both a successful and complementary method to screen for nearby interacting proteins of HIV-1 Gag during the replicative cycle in different cell lines. * Correspondence: andrew.mouland@mcgill.ca 1HIV-1 RNA Trafficking Laboratory, Lady Davis Institute at the Jewish General Hospital, Montréal, Québec H3T 1E2, Canada 2Department of Medicine, McGill University, Montréal, Québec H3A 0G4, Canada Full list of author information is available at the end of the article

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