Detection of human immunodeficiency virus type 1 Nef and CD4 physical interaction in living human cells by using bioluminescence resonance energy transfer.
CD4 down-regulation by human immunodeficiency virus type 1 (HIV-1) Nef protein is a key function for virus virulence. This activity may be mediated by a direct Nef-CD4 interaction. We investigated the formation, in situ, of such a complex between proteins using bioluminescence resonance energy transfer technology and co-immunoprecipitations. Our data clearly demonstrate that Nef and CD4 interact in intact human cells. Moreover, our results clearly indicate that the dileucine motif of the CD4 cytoplasmic domain, critical for the Nef-induced CD4 down-regulation, is not implicated in the Nef/CD4 complex formation in the cellular context.
Cluet
David
D
INSERM-ULP U544, Université Louis Pasteur, Institut de Virologie, 3 Rue Koeberlé, 67000 Strasbourg, France.
Bertsch
Christophe
C
Beyer
Christian
C
Gloeckler
Liliane
L
Erhardt
Mathieu
M
Gut
Jean-Pierre
JP
Galzi
Jean-Luc
JL
Aubertin
Anne-Marie
AM
D015703:Antigens, CD / Q000276:immunology; D001426:Bacterial Proteins / Q000032:analysis; D001665:Binding Sites; D015704:CD4 Antigens / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000821:virology*; D002460:Cell Line; D015693:Gene Products, nef / Q000276:immunology*; D017930:Genes, Reporter; D015497:HIV-1 / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D006801:Humans; D007668:Kidney; D008163:Luminescent Measurements / Q000379:methods; D008164:Luminescent Proteins / Q000032:analysis; D054311:nef Gene Products, Human Immunodeficiency Virus
D015703:Antigens, CD; D001426:Bacterial Proteins; D015704:CD4 Antigens; D015693:Gene Products, nef; D008164:Luminescent Proteins; D054311:nef Gene Products, Human Immunodeficiency Virus; C054562:yellow fluorescent protein, Bacteria