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Open Access Highly Accessed Research

Characterization of the HIV-1 RNA associated proteome identifies Matrin 3 as a nuclear cofactor of Rev function

Anna Kula1, Jessica Guerra13, Anna Knezevich1, Danijela Kleva1, Michael P Myers2 and Alessandro Marcello1*

Author Affiliations

1 Laboratory of Molecular Virology, International Centre for Genetic Engineering and Biotechnology (ICGEB), Padriciano, 99, 34012 Trieste, Italy

2 Laboratory of Protein Networks, International Centre for Genetic Engineering and Biotechnology (ICGEB), Padriciano, 99, 34012 Trieste, Italy

3 Department of Microbiology and Molecular Medicine, University of Geneva, Geneva, 1211, Switzerland

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Retrovirology 2011, 8:60  doi:10.1186/1742-4690-8-60

Published: 20 July 2011

Abstract

Background

Central to the fully competent replication cycle of the human immunodeficiency virus type 1 (HIV-1) is the nuclear export of unspliced and partially spliced RNAs mediated by the Rev posttranscriptional activator and the Rev response element (RRE).

Results

Here, we introduce a novel method to explore the proteome associated with the nuclear HIV-1 RNAs. At the core of the method is the generation of cell lines harboring an integrated provirus carrying RNA binding sites for the MS2 bacteriophage protein. Flag-tagged MS2 is then used for affinity purification of the viral RNA. By this approach we found that the viral RNA is associated with the host nuclear matrix component MATR3 (Matrin 3) and that its modulation affected Rev activity. Knockdown of MATR3 suppressed Rev/RRE function in the export of unspliced HIV-1 RNAs. However, MATR3 was able to associate with Rev only through the presence of RRE-containing viral RNA.

Conclusions

In this work, we exploited a novel proteomic method to identify MATR3 as a cellular cofactor of Rev activity. MATR3 binds viral RNA and is required for the Rev/RRE mediated nuclear export of unspliced HIV-1 RNAs.