P2Y1 receptor signaling is controlled by interaction with the PDZ scaffold NHERF-2.
P2Y(1) purinergic receptors (P2Y(1)Rs) mediate rises in intracellular Ca(2+) in response to ATP, but the duration and characteristics of this Ca(2+) response are known to vary markedly in distinct cell types. We screened the P2Y(1)R carboxyl terminus against a recently created proteomic array of PDZ (PSD-95/Drosophila Discs large/ZO-1 homology) domains and identified a previously unrecognized, specific interaction with the second PDZ domain of the scaffold NHERF-2 (Na(+)/H(+) exchanger regulatory factor type 2). Furthermore, we found that P2Y(1)R and NHERF-2 associate in cells, allowing NHERF-2-mediated tethering of P2Y(1)R to key downstream effectors such as phospholipase Cbeta. Finally, we found that coexpression of P2Y(1)R with NHERF-2 in glial cells prolongs P2Y(1)R-mediated Ca(2+) signaling, whereas disruption of the P2Y(1)R-NHERF-2 interaction by point mutations attenuates the duration of P2Y(1)R-mediated Ca(2+) responses. These findings reveal that NHERF-2 is a key regulator of the cellular activity of P2Y(1)R and may therefore determine cell-specific differences in P2Y(1)R-mediated signaling.
Fam
Sami R
SR
Department of Pharmacology, Yerkes National Primate Research Center, and Division of Digestive Disease, Emory University School of Medicine, Atlanta, GA 30322, USA.
Paquet
Maryse
M
Castleberry
Amanda M
AM
Oller
Heide
H
Lee
C Justin
CJ
Traynelis
Stephen F
SF
Smith
Yoland
Y
Yun
C Chris
CC
Hall
Randy A
RA
D015153:Blotting, Western; D001921:Brain / Q000378:metabolism / Q000648:ultrastructure; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D008854:Microscopy, Electron; D009457:Neuroglia / Q000378:metabolism / Q000648:ultrastructure; D010750:Phosphoproteins; D040081:Protein Array Analysis; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D018048:Receptors, Purinergic P2 / Q000378:metabolism*; D058472:Receptors, Purinergic P2Y1; D015398:Signal Transduction / Q000502:physiology*; D017923:Sodium-Hydrogen Exchangers; D014162:Transfection
D003598:Cytoskeletal Proteins; C546382:P2RY1 protein, human; D010750:Phosphoproteins; D018048:Receptors, Purinergic P2; D058472:Receptors, Purinergic P2Y1; D017923:Sodium-Hydrogen Exchangers; C093376:sodium-hydrogen exchanger regulatory factor; D002118:Calcium