Definition of the interacting interfaces of Apobec3G and HIV-1 Vif using MAPPIT mutagenesis analysis

Nucleic Acids Res. 2010 Apr;38(6):1902-12. doi: 10.1093/nar/gkp1154. Epub 2009 Dec 16.

Abstract

The host restriction factor Apobec3G is a cytidine deaminase that incorporates into HIV-1 virions and interferes with viral replication. The HIV-1 accessory protein Vif subverts Apobec3G by targeting it for proteasomal degradation. We propose a model in which Apobec3G N-terminal domains symmetrically interact via a head-to-head interface containing residues 122 RLYYFW 127. To validate this model and to characterize the Apobec3G-Apobec3G and the Apobec3G-Vif interactions, the mammalian protein-protein interaction trap two-hybrid technique was used. Mutations in the head-to-head interface abrogate the Apobec3G-Apobec3G interaction. All mutations that inhibit Apobec3G-Apobec3G binding also inhibit the Apobec3G-Vif interaction, indicating that the head-to head interface plays an important role in the interaction with Vif. Only the D128K, P129A and T32Q mutations specifically affect the Apobec3G-Vif association. In our model, D128, P129 and T32 cluster at the edge of the head-to-head interface, possibly forming a Vif binding site composed of two Apobec3G molecules. We propose that Vif either binds at the Apobec3G head-to-head interface or associates with an RNA-stabilized Apobec3G oligomer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • APOBEC-3G Deaminase
  • Binding Sites
  • Cell Line
  • Cytidine Deaminase / chemistry*
  • Cytidine Deaminase / genetics
  • Cytidine Deaminase / metabolism
  • Cytosine Deaminase / chemistry
  • Dimerization
  • Humans
  • Models, Molecular
  • Mutagenesis, Site-Directed
  • Protein Interaction Domains and Motifs
  • Structural Homology, Protein
  • Two-Hybrid System Techniques
  • vif Gene Products, Human Immunodeficiency Virus / chemistry*
  • vif Gene Products, Human Immunodeficiency Virus / metabolism

Substances

  • vif Gene Products, Human Immunodeficiency Virus
  • vif protein, Human immunodeficiency virus 1
  • APOBEC3F protein, human
  • Cytosine Deaminase
  • APOBEC-3G Deaminase
  • APOBEC3G protein, human
  • Cytidine Deaminase