Rab5a and rab11a mediate agonist-induced trafficking of protease-activated receptor 2.
We evaluated the contribution of rab5a and rab11a to trafficking and signaling of protease-activated receptor 2 (PAR2), a receptor for trypsin and tryptase. Agonists stimulated internalization of PAR2 into early endosomes containing rab5a. Dominant negative rab5aS34N disrupted early endosomes and inhibited agonist-stimulated endocytosis of PAR2. Internalized PAR2 was sorted to lysosomes, and rab5a remained in early endosomes. Rab5a promoted and rab5aS34N impeded resensitization of trypsin-induced calcium mobilization. Rab11a was detected in the Golgi apparatus with PAR2, and PAR2 agonists stimulated redistribution of rab11a into vesicles containing PAR2 that migrated to the cell surface. Dominant negative rab11aS25N was mostly confined to the Golgi apparatus. Although expression of rab11aS25N caused retention of PAR2 in the Golgi apparatus, it did not abolish trafficking of PAR2 to the cell surface. However, expression of wild-type rab11a accelerated both recovery of PAR2 at the cell surface and resensitization of PAR2 signaling. Thus rab5a is required for PAR2 endocytosis and resensitization, whereas rab11a contributes to trafficking of PAR2 from the Golgi apparatus to the plasma membrane.
Roosterman
Dirk
D
Department of Surgery, University of California, San Francisco, California 94143-0660, USA.
Schmidlin
Fabien
F
Bunnett
Nigel W
NW
D000818:Animals; D002460:Cell Line; D004705:Endocytosis / Q000502:physiology; D006801:Humans; D021381:Protein Transport / Q000502:physiology; D051381:Rats; D044464:Receptor, PAR-2; D018179:Receptors, Thrombin / Q000819:agonists* / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D014018:Tissue Distribution; D020691:rab GTP-Binding Proteins / Q000502:physiology*; D020696:rab5 GTP-Binding Proteins / Q000502:physiology*; D000099193:rab11 GTP-Binding Proteins
D044464:Receptor, PAR-2; D018179:Receptors, Thrombin; D020691:rab GTP-Binding Proteins; D020696:rab5 GTP-Binding Proteins; D000099193:rab11 GTP-Binding Proteins