Palmitoylation-dependent control of degradation, life span, and membrane expression of the CCR5 receptor.
We have shown that the chemokine and HIV receptor CCR5 is palmitoylated on a cluster of cysteine residues located at the boundary between the seventh transmembrane region and the cytoplasmic tail. Single or combined substitutions of the three cysteines (Cys-321, Cys-323, and Cys-324) or incubation of wild-type CCR5-transfected cells with the palmitic acid analog 2-bromopalmitate prevented palmitoylation of the receptor. Moreover, failure of CCR5 to be palmitoylated resulted in both accumulation in intracellular stores and a profound decrease of membrane expression of the receptor. Upon metabolic labeling, kinetic experiments showed that the half-life of palmitoylation-deficient CCR5 is profoundly decreased. Bafilomycin A1, but not a specific proteasome inhibitor, prevented early degradation of palmitoylation-deficient CCR5 and promoted its accumulation in lysosomal compartments. Although membrane expression of the CCR5 mutant was diminished, the molecules reaching the membrane were still able to interact efficiently with the chemokine ligand MIP1 beta and remained able to function as HIV co-receptors. Thus we conclude that palmitoylation controls CCR5 expression through regulation of the life span of this receptor.
Percherancier
Y
Y
Institut Pasteur, Unité d'Immunologie Virale, 25-28 rue du Dr. Roux, 75724 Paris, cedex 15, France.
Planchenault
T
T
Valenzuela-Fernandez
A
A
Virelizier
J L
JL
Arenzana-Seisdedos
F
F
Bachelerie
F
F
D000595:Amino Acid Sequence; D002460:Cell Line; D003545:Cysteine / Q000378:metabolism; D005434:Flow Cytometry; D006207:Half-Life; D006801:Humans; D006868:Hydrolysis; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D019308:Palmitic Acid / Q000378:metabolism*; D019713:Receptors, CCR5 / Q000737:chemistry / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
D008565:Membrane Proteins; D019713:Receptors, CCR5; D019308:Palmitic Acid; D003545:Cysteine