MassIVE MSV000079605

Partial Public PXD003861

MudPIT analysis of HIV-1 Vpr-asssociated proteins from the permissive CEM.SS human T4-lymphoblastoid cell line

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Description

CEM.SS-iH1.Vpr cells expressing a tandem HA-FLAG- epitope-tagged HIV-1 NL4-3 Vpr (hfa-H.Vpr) under control of the Lenti-X Tet-On 3G Inducible Expression System (Clontech), and control CEM.SS cells (3x105 cells/ml, 4 liters), were induced with doxycycline (0.1ug/ml) for 9 hours. Whole cell extracts were prepared and tandem affinity purifications of Vpr and its associated proteins, from 5 grams of wet cell mass, were performed as described (Hrecka K, et al. Nature 2011; 474(7353):658-661). The Vpr-protein complexes and negative controls purified from T cells were digested with endoproteinase LysC followed by trypsin and the resulting peptide mixtures were analyzed by Multidimensional Protein Identification Technology (MudPIT) on an LTQ ion trap mass spectrometer as described (Florens L, Washburn MP. Methods Mol Biol 2006;328:159-75). [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: HIV accessory virulence protein Vpr ; affinity purification ; T cells

Contact

Principal Investigators:
(in alphabetical order) 		Laurence Florens
Submitting User: laflorens

Publications

Hrecka K, Hao C, Shun MC, Kaur S, Swanson SK, Florens L, Washburn MP, Skowronski J.
HIV-1 and HIV-2 exhibit divergent interactions with HLTF and UNG2 DNA repair proteins.
Proc. Natl. Acad. Sci. U.S.A. Epub 2016 Jun 22.

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Originally identified proteins that were automatically remapped by MassIVE to proteins in the SwissProt human reference database.

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Number of distinct proteins found to be differentially abundant in at least one comparison across all analyses (original submission and reanalyses) associated with this dataset.

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