Host cell interactome of HIV-1 Rev includes RNA helicases involved in multiple facets of virus production
| Autor: | Andrew C. S. Saphire, Souad Naji, Géza Ambrus, Paul Vesely, Peter Cimermancic, Michael D. Huber, Jason R. Reyes, Nevan J. Krogan, Jeffrey R. Johnson, John R. Yates, Larry Gerace, Rebecca Filbrandt |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Proteomics
viruses HIV Infections Virus Replication Biochemistry Interactome Virus Analytical Chemistry DEAD-box RNA Helicases chemistry.chemical_compound Escherichia coli Humans RNA Small Interfering Nuclear export signal Molecular Biology Genetics DDX5 biology Research Helicase RNA rev Gene Products Human Immunodeficiency Virus Viral replication chemistry Host-Pathogen Interactions biology.protein HIV-1 HeLa Cells |
| Zdroj: | Molecularcellular proteomics : MCP. 11(4) |
| ISSN: | 1535-9484 |
| Popis: | The HIV-1 Rev protein plays a key role in the late phase of virus replication. It binds to the Rev Response Element found in underspliced HIV mRNAs, and drives their nuclear export by the CRM1 receptor pathway. Moreover, mounting evidence suggests that Rev has additional functions in viral replication. Here we employed proteomics and statistical analysis to identify candidate host cell factors that interact with Rev. For this we studied Rev complexes assembled in vitro with nuclear or cytosolic extracts under conditions emulating various intracellular environments of Rev. We ranked the protein-protein interactions by combining several statistical features derived from pairwise comparison of conditions in which the abundance of the binding partners changed. As a validation set, we selected the eight DEAD/H box proteins of the RNA helicase family from the top-ranking 5% of the proteins. These proteins all associate with ectopically expressed Rev in immunoprecipitates of cultured cells. From gene knockdown approaches, our work in combination with previous studies indicates that six of the eight DEAD/H proteins are linked to HIV production in our cell model. In a more detailed analysis of infected cells where either DDX3X, DDX5, DDX17, or DDX21 was silenced, we observed distinctive phenotypes for multiple replication features, variously involving virus particle release, the levels of unspliced and spliced HIV mRNAs, and the nuclear and cytoplasmic concentrations of these transcripts. Altogether the work indicates that our top-scoring data set is enriched in Rev-interacting proteins relevant to HIV replication. Our more detailed analysis of several Rev-interacting DEAD proteins suggests a complex set of functions for the helicases in regulation of HIV mRNAs. The strategy used here for identifying Rev interaction partners should prove effective for analyzing other viral and cellular proteins. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|