Brain transcriptome-wide screen for HIV-1 Nef protein interaction partners reveals various membrane-associated proteins

PLoS One. 2012;7(12):e51578. doi: 10.1371/journal.pone.0051578. Epub 2012 Dec 17.

Abstract

HIV-1 Nef protein contributes essentially to the pathology of AIDS by a variety of protein-protein-interactions within the host cell. The versatile functionality of Nef is partially attributed to different conformational states and posttranslational modifications, such as myristoylation. Up to now, many interaction partners of Nef have been identified using classical yeast two-hybrid screens. Such screens rely on transcriptional activation of reporter genes in the nucleus to detect interactions. Thus, the identification of Nef interaction partners that are integral membrane proteins, membrane-associated proteins or other proteins that do not translocate into the nucleus is hampered. In the present study, a split-ubiquitin based yeast two-hybrid screen was used to identify novel membrane-localized interaction partners of Nef. More than 80% of the hereby identified interaction partners of Nef are transmembrane proteins. The identified hits are GPM6B, GPM6A, BAP31, TSPAN7, CYB5B, CD320/TCblR, VSIG4, PMEPA1, OCIAD1, ITGB1, CHN1, PH4, CLDN10, HSPA9, APR-3, PEBP1 and B3GNT, which are involved in diverse cellular processes like signaling, apoptosis, neurogenesis, cell adhesion and protein trafficking or quality control. For a subfraction of the hereby identified proteins we present data supporting their direct interaction with HIV-1 Nef. We discuss the results with respect to many phenotypes observed in HIV infected cells and patients. The identified Nef interaction partners may help to further elucidate the molecular basis of HIV-related diseases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Biomarkers / metabolism
  • Blotting, Western
  • Brain / metabolism*
  • COS Cells
  • Chlorocebus aethiops
  • Humans
  • Immunoenzyme Techniques
  • Immunoprecipitation
  • Membrane Proteins / antagonists & inhibitors
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Molecular Sequence Data
  • Protein Binding
  • Protein Interaction Mapping*
  • RNA, Small Interfering / genetics
  • Sequence Homology, Amino Acid
  • Transcriptome*
  • Two-Hybrid System Techniques
  • Ubiquitin / metabolism
  • nef Gene Products, Human Immunodeficiency Virus / genetics
  • nef Gene Products, Human Immunodeficiency Virus / metabolism*

Substances

  • Biomarkers
  • Membrane Proteins
  • RNA, Small Interfering
  • Ubiquitin
  • nef Gene Products, Human Immunodeficiency Virus
  • nef protein, Human immunodeficiency virus 1

Grant support

This work was supported by a grant from the Präsidentenfond der Helmholtzgemeinschaft (HGF, Virtual Institute of Structural Biology) to DW. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.