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

Edward J D Greenwood; James C Williamson; Agata Sienkiewicz; Adi Naamati; Nicholas J Matheson; Paul J Lehner

doi:10.1016/j.celrep.2019.04.025

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

Cell Rep. 2019 Apr 30;27(5):1579-1596.e7. doi: 10.1016/j.celrep.2019.04.025.

Authors

Edward J D Greenwood¹, James C Williamson², Agata Sienkiewicz³, Adi Naamati⁴, Nicholas J Matheson⁴, Paul J Lehner⁵

Affiliations

¹ Department of Medicine, Cambridge Biomedical Campus, University of Cambridge, Cambridge CB2 0QQ, UK; Cambridge Institute for Medical Research, Keith Peters Building, Cambridge Biomedical Campus, University of Cambridge, Cambridge CB2 0XY, UK; Cambridge Institute of Therapeutic Immunology and Infectious Disease, Jeffrey Cheah Biomedical Centre, Cambridge Biomedical Campus, University of Cambridge, Cambridge CB2 0AW, UK. Electronic address: ejdg2@cam.ac.uk.
² Department of Medicine, Cambridge Biomedical Campus, University of Cambridge, Cambridge CB2 0QQ, UK; Cambridge Institute for Medical Research, Keith Peters Building, Cambridge Biomedical Campus, University of Cambridge, Cambridge CB2 0XY, UK; Cambridge Institute of Therapeutic Immunology and Infectious Disease, Jeffrey Cheah Biomedical Centre, Cambridge Biomedical Campus, University of Cambridge, Cambridge CB2 0AW, UK. Electronic address: jcw76@cam.ac.uk.
³ Department of Medicine, Cambridge Biomedical Campus, University of Cambridge, Cambridge CB2 0QQ, UK; Cambridge Institute for Medical Research, Keith Peters Building, Cambridge Biomedical Campus, University of Cambridge, Cambridge CB2 0XY, UK; Cambridge Institute of Therapeutic Immunology and Infectious Disease, Jeffrey Cheah Biomedical Centre, Cambridge Biomedical Campus, University of Cambridge, Cambridge CB2 0AW, UK.
⁴ Department of Medicine, Cambridge Biomedical Campus, University of Cambridge, Cambridge CB2 0QQ, UK; Cambridge Institute of Therapeutic Immunology and Infectious Disease, Jeffrey Cheah Biomedical Centre, Cambridge Biomedical Campus, University of Cambridge, Cambridge CB2 0AW, UK.
⁵ Department of Medicine, Cambridge Biomedical Campus, University of Cambridge, Cambridge CB2 0QQ, UK; Cambridge Institute for Medical Research, Keith Peters Building, Cambridge Biomedical Campus, University of Cambridge, Cambridge CB2 0XY, UK; Cambridge Institute of Therapeutic Immunology and Infectious Disease, Jeffrey Cheah Biomedical Centre, Cambridge Biomedical Campus, University of Cambridge, Cambridge CB2 0AW, UK. Electronic address: pjl30@cam.ac.uk.

Abstract

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 demonstrate that Vpr is both necessary and sufficient for the DCAF1/DDB1/CUL4 E3 ubiquitin ligase-mediated degradation of at least 38 cellular proteins, causing systems-level changes to the cellular proteome. We therefore propose that promiscuous targeting of multiple host factors underpins complex Vpr-dependent cellular phenotypes and validate this in the case of G2/M cell cycle arrest. Our model explains how Vpr modulates so many cell biological processes and why the functional consequences of previously described Vpr targets, identified and studied in isolation, have proved elusive.

Keywords: HIV; IP-MS; SIV; TMT; Vpr; Vpx; mass spectrometry; proteomics; pulsed SILAC.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line, Tumor
Conserved Sequence
G2 Phase Cell Cycle Checkpoints
HEK293 Cells
HIV Infections / metabolism*
HIV Infections / virology
HIV-1 / metabolism*
HIV-1 / pathogenicity
Humans
Proteolysis
Proteome / chemistry
Proteome / metabolism*
Ubiquitin-Protein Ligases / metabolism
Ubiquitination
vpr Gene Products, Human Immunodeficiency Virus / genetics
vpr Gene Products, Human Immunodeficiency Virus / metabolism*

Substances

Proteome
vpr Gene Products, Human Immunodeficiency Virus
Ubiquitin-Protein Ligases

Abstract

Publication types

MeSH terms

Substances

Grants and funding