Dataset Information

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

ABSTRACT: 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).

INSTRUMENT(S): LTQ

ORGANISM(S): Hiv-1 Vector Pnl4-3 (ncbitaxon:151458) Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Laurence Florens

PROVIDER: MSV000079605 | MassIVE | Thu Mar 24 11:22:00 GMT 2016

SECONDARY ACCESSION(S): PXD003861

REPOSITORIES: MassIVE

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Publications

HIV-1 and HIV-2 exhibit divergent interactions with HLTF and UNG2 DNA repair proteins.

Hrecka Kasia K Hao Caili C Shun Ming-Chieh MC Kaur Sarabpreet S Swanson Selene K SK Florens Laurence L Washburn Michael P MP Skowronski Jacek J

Proceedings of the National Academy of Sciences of the United States of America 20160622 27

HIV replication in nondividing host cells occurs in the presence of high concentrations of noncanonical dUTP, apolipoprotein B mRNA-editing, enzyme-catalytic, polypeptide-like 3 (APOBEC3) cytidine deaminases, and SAMHD1 (a cell cycle-regulated dNTP triphosphohydrolase) dNTPase, which maintains low concentrations of canonical dNTPs in these cells. These conditions favor the introduction of marks of DNA damage into viral cDNA, and thereby prime it for processing by DNA repair enzymes. Accessory pr ...[more]

PMID: 27335459

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