Dataset Information

Massive Proteome Remodelling by HIV Vpr

ABSTRACT: Viruses target host proteins for degradation to enhance their replication and transmission, and identifying these targets has provided key insights into the host-pathogen interaction1-3. Here, we use complementary unbiased mass spectrometry-based approaches to dissect the widespread proteomeic remodelling seen in HIV-1 infected T-cells. Remarkably, the HIV accessory protein Vpr is both necessary and sufficient to cause the vast majority of these changes. Protein regulation requires recruitment of the DCAF1/DDB1/CUL4 E3 ubiquitin ligase complex, and pulsed-Stable Isotope Labelling with Amino Acids in Cell Culture (SILAC) and immunoprecipitation-mass spectrometry (IP-MS) identified at least 38 cellular proteins directly targeted for degradation by Vpr. Whilst other HIV-1 accessory proteins downregulate a small number of specific host factors, Vpr depletes multiple protein targets, causing systems-level changes to the cellular proteome. A subset of the novel cellular targets identified in this study are depleted by Vpr variants from across HIV-1/SIVcpz and other primate lentiviral lineages, confirming their biological importance in vivo.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): T Cell

DISEASE(S): Human Immunodeficiency Virus Infectious Disease

SUBMITTER: James Williamson

LAB HEAD: Paul Lehner

PROVIDER: PXD013221 | Pride | 2019-05-01

REPOSITORIES: Pride

ACCESS DATA

Publications

Promiscuous Targeting of Cellular Proteins by Vpr Drives Systems-Level Proteomic Remodeling in HIV-1 Infection.

Greenwood Edward J D EJD Williamson James C JC Sienkiewicz Agata A Naamati Adi A Matheson Nicholas J NJ Lehner Paul J PJ

Cell reports 20190401 5

HIV-1 encodes four "accessory proteins" (Vif, Vpr, Vpu, and Nef), dispensable for viral replication in vitro but essential for viral pathogenesis in vivo. Well characterized cellular targets have been associated with Vif, Vpu, and Nef, which counteract host restriction and promote viral replication. Conversely, although several substrates of Vpr have been described, their biological significance remains unclear. Here, we use complementary unbiased mass spectrometry-based approaches to demonstrat ...[more]

PMID: 31042482

Similar Datasets

Project description:Viruses have an ambivalent relationship with the host proteome. While several host proteins are indispensable for replication, detrimental restriction factors need to be antagonized. This process is frequently catalysed by viral accessory proteins which exploit the ubiquitin proteasome pathway, DDB1, and Cullin Ubiquitin ligases for the degradation of restriction factors, like in the case of the mouse cytomegalovirus encoded interferon antagonist pM27 which exploits DDB1 for STAT2 degradation. pM27 homologous proteins are conserved in betaherpesviruses. DDB1 and Cullins also serve as essential co-factor for accessory proteins encoded by other virus families like Vpr (HIV-1- and -2-encoded), Vpx (HIV-2-encoded), HBx (HBV-encoded) and its woodchuck hepatitis virus homolog (WHx). To establish a comprehensive understanding of the complexes assembled by viral DDB1-interacting proteins and their targets, we performed immuno-affinity precipitations for a panel of viral proteins known or suspected to interact with DDB1 and/or Cullins. For each protein, we conducted four biological replicates and identified and quantified all co-precipitating proteins by mass spectrometry. Empty plasmids and the HCMV-encoded protein pUL42, which binds the unrelated ubiquitin ligase Itch, served as negative controls and outside group, respectively. This analysis confirmed previously described interactions (pM27 with STAT2), extended known interactions to paralogous proteins (e.g. HBx and WHx with different Spindlin proteins), and documented anticipated interactions for the first time (e.g. Vpr with Bax). The comparison revealed considerable differences concerning the interaction with DDB1 and Cullin 4A/B in terms of the general ability as well as the strength of interaction. On global level, the correlation of the interactomes fitted well with the phylogenetic relationship of the proteins but also highlighted a considerable overlap between the DDB1 interactors of non-related viruses (e.g. herpesviral and hepadnaviral).

Dataset Information

Massive Proteome Remodelling by HIV Vpr

Publications

Promiscuous Targeting of Cellular Proteins by Vpr Drives Systems-Level Proteomic Remodeling in HIV-1 Infection.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure