The human polycomb group EED protein interacts with the integrase of human immunodeficiency virus type 1.

Sébastien Violot (1) , Saw See Hong (1) , Dina Rakotobe (1) , Caroline Petit (2) , Bernard Gay (1) , Karen Moreau (1) , Geneviève Billaud (1) , Stéphane Priet (3) , Joséphine Sire (3) , Olivier Schwartz (2) , Jean-François Mouscadet (4) , Pierre Boulanger (1)

1 VPV - Virologie et pathogenèse virale
2 Rétrovirus et Transfert Génétique
3 Pathogénie des infections à lentivirus
4 LBPA - Laboratoire de Biotechnologie et Pharmacogénétique Appliquée

Human EED, a member of the superfamily of WD-40 repeat proteins and of the Polycomb group proteins, has been identified as a cellular partner of the human immunodeficiency virus type 1 (HIV-1) matrix (MA) protein (R. Peytavi et al., J. Biol. Chem. 274:1635-1645, 1999). In the present study, EED was found to interact with HIV-1 integrase (IN) both in vitro and in vivo in yeast. In vitro, data from mutagenesis studies, pull-down assays, and phage biopanning suggested that EED-binding site(s) are located in the C-terminal domain of IN, between residues 212 and 264. In EED, two putative discrete IN-binding sites were mapped to its N-terminal moiety, at a distance from the MA-binding site, but EED-IN interaction also required the integrity of the EED last two WD repeats. EED showed an apparent positive effect on IN-mediated DNA integration reaction in vitro, in a dose-dependent manner. In situ analysis by immunoelectron microscopy (IEM) of cellular distribution of IN and EED in HIV-1-infected cells (HeLa CD4(+) cells or MT4 lymphoid cells) showed that IN and EED colocalized in the nucleus and near nuclear pores, with maximum colocalization events occurring at 6 h postinfection (p.i.). Triple colocalizations of IN, EED, and MA were also observed in the nucleoplasm of infected cells at 6 h p.i., suggesting the ocurrence of multiprotein complexes involving these three proteins at early steps of the HIV-1 virus life cycle. Such IEM patterns were not observed with a noninfectious, envelope deletion mutant of HIV-1.

Mots clés: Non-U.S. Gov't; Repressor Proteins/genetics/*metabolism/ultrastructure; Research Support; Sequence Homology; Amino Acid; Two-Hybrid System Techniques; Recombinant Proteins/genetics/metabolism/ultrastructure; Site-Directed; Mutagenesis; Protein Binding; Molecular Sequence Data; Immunoelectron; Amino Acid Sequence; Microscopy; Humans; HIV-1/enzymology/metabolism/physiology; HIV Integrase/*metabolism; Cell Line

Domaines: Virologie

Sébastien Violot, Saw See Hong, Dina Rakotobe, Caroline Petit, Bernard Gay, et al.. The human polycomb group EED protein interacts with the integrase of human immunodeficiency virus type 1.. Journal of Virology, 2003, 77 (23), pp.12507-22. ⟨10.1128/JVI.77.23.12507-12522.2003⟩. ⟨hal-00119648⟩