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

被引:55
作者
Violot, S
Hong, SS
Rakotobe, D
Petit, C
Gay, B
Moreau, K
Billaud, G
Priet, S
Sire, J
Schwartz, O
Mouscadet, JF
Boulanger, P
机构
[1] RTH Laennec Lyon, Fac Med, Lab Virol & Pathogenese Virale, CNRS,UMR 5537, F-69372 Lyon 08, France
[2] Univ Lyon 1, F-69372 Lyon 08, France
[3] Inst Pasteur, Lab Retrovirus & Transfert Genet, F-75724 Paris 15, France
[4] INSERM, U372, Unite Pathogenie Infect Lentivirus, F-13276 Marseille 09, France
[5] Ecole Normale Super, CNRS, UMR 8532, F-75235 Cachan, France
关键词
D O I
10.1128/JVI.77.23.12507-12522.2003
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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.
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页码:12507 / 12522
页数:16
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