Pathways for internalization and recycling of the chemokine receptor CCR5.
M-tropic human immunodeficiency virus (HIV-1) strains enter the cell after interaction with their receptors, CD4 and the G-protein-coupled chemokine receptor CCR5. The number of cell surface CCR5 molecules is thought to be important in determining the infection rate for HIV. Cell surface CCR5 is dependent on the rate of receptor internalization and recycling. Internalization of G-protein-coupled receptors after agonist activation is thought to occur either through clathrin-coated pits or through caveolae. In this study, the role of these different pathways was investigated in Chinese hamster ovary cells expressing CCR5 using specific inhibitors. Internalization of CCR5 after chemokine treatment was inhibited by sucrose, indicating a role for the clathrin-coated pit pathway. Activation of CCR5 leads to arrestin-2 movement in the cells, providing further evidence for the involvement of clathrin-coated pits. Nystatin and filipin also affected the rate of internalization of CCR5, indicating a role for caveolae. Using inhibitors of vesicle transport in the cell, it was found that the CCR5 recycling pathway is independent of the Golgi apparatus and late endosomes. Protein synthesis is not involved in receptor recovery. It seems likely that after internalization, CCR5 is directed to early endosomes and subsequently recycled to the cell surface.
Mueller
Anja
A
School of Animal and Microbial Sciences, University of Reading, Reading, RG6 6AJ, United Kingdom.
Kelly
Eamonn
E
Strange
Philip G
PG
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D016466:CHO Cells; D021941:Caveolae; D022163:Clathrin-Coated Vesicles / Q000502:physiology; D006224:Cricetinae; D004705:Endocytosis / Q000187:drug effects; D005372:Filipin / Q000494:pharmacology; D018613:Microscopy, Confocal; D009761:Nystatin / Q000494:pharmacology; D015388:Organelles; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D021381:Protein Transport / Q000187:drug effects; D019713:Receptors, CCR5 / Q000378:metabolism* / Q000502:physiology; D013395:Sucrose / Q000494:pharmacology; D014162:Transfection
D000900:Anti-Bacterial Agents; D011500:Protein Synthesis Inhibitors; D019713:Receptors, CCR5; D009761:Nystatin; D013395:Sucrose; D005372:Filipin