Interaction between the cytoplasmic domains of HIV-1 Vpu and CD4: role of Vpu residues involved in CD4 interaction and in vitro CD4 degradation.
The Vpu and CD4 cytoplasmic domains were found, by using a two-hybrid assay in yeast, to interact in the absence of their membrane anchor domains. Studies on several deletion and point mutants revealed that the overall structure of the Vpu cytoplasmic domain is required for this interaction. The Vpu amino acid residues involved in the interaction with CD4 were identified. Deletion of the C-terminal residues of Vpu, required for CD4 degradation, as well as the double mutation on the casein kinase II phosphorylation sites S52N-S56N, also involved in CD4 degradation, resulted in the loss of interaction with CD4 and in the inability to induce CD4 degradation. These results suggest that the ability of Vpu to mediate the degradation of CD4 is linked to its capacity to physically interact with CD4. However, additional mutagenesis on the S52 site revealed that the interaction between the cytoplasmic domains of Vpu and CD4 is not sufficient for in vitro Vpu-mediated CD4 degradation.
Margottin
F
F
Unité 332, INSERM, ICGM Institut Cochin de Génétique moléculaire, Université Paris V, France.
Benichou
S
S
Durand
H
H
Richard
V
V
Liu
L X
LX
Gomas
E
E
Benarous
R
R
D000595:Amino Acid Sequence; D015704:CD4 Antigens / Q000235:genetics* / Q000378:metabolism* / Q000502:physiology; D047390:Casein Kinase II; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism*; D054298:Human Immunodeficiency Virus Proteins; D008969:Molecular Sequence Data; D017354:Point Mutation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D054334:Viral Regulatory and Accessory Proteins / Q000235:genetics* / Q000378:metabolism* / Q000502:physiology; D015003:Yeasts; D000086082:Viroporin Proteins
D015704:CD4 Antigens; D047390:Casein Kinase II; D054298:Human Immunodeficiency Virus Proteins; D017346:Protein Serine-Threonine Kinases; D054334:Viral Regulatory and Accessory Proteins; C514859:vpu protein, Human immunodeficiency virus 1; D000086082:Viroporin Proteins