Substance P-induced trafficking of beta-arrestins. The role of beta-arrestins in endocytosis of the neurokinin-1 receptor.
Agonist-induced redistribution of G-protein-coupled receptors (GPCRs) and beta-arrestins determines the subsequent cellular responsiveness to agonists and is important for signal transduction. We examined substance P (SP)-induced trafficking of beta-arrestin1 and the neurokinin-1 receptor (NK1R) in KNRK cells in real time using green fluorescent protein. Green fluorescent protein did not alter function or localization of the NK1R or beta-arrestin1. SP induced (a) striking and rapid (<1 min) translocation of beta-arrestin1 from the cytosol to the plasma membrane, which preceded NK1R endocytosis; (b) redistribution of the NK1R and beta-arrestin1 into the same endosomes containing SP and the transferrin receptor (2-10 min); (c) prolonged colocalization of the NK1R and beta-arrestin1 in endosomes (>60 min); (d) gradual resumption of the steady state distribution of the NK1R at the plasma membrane and beta-arrestin1 in the cytosol (4-6 h). SP stimulated a similar redistribution of immunoreactive beta-arrestin1 and beta-arrestin2. In contrast, SP did not affect Galphaq/11 distribution, which remained at the plasma membrane. Expression of the dominant negative beta-arrestin319-418 inhibited SP-induced endocytosis of the NK1R. Thus, SP induces rapid translocation of beta-arrestins to the plasma membrane, where they participate in NK1R endocytosis. beta-Arrestins colocalize with the NK1R in endosomes until the NK1R recycles and beta-arrestins return to the cytosol.
McConalogue
K
K
Department of Surgery, University of California, San Francisco, California 94143-0660, USA.
Déry
O
O
Lovett
M
M
Wong
H
H
Walsh
J H
JH
Grady
E F
EF
Bunnett
N W
NW
D000818:Animals; D019390:Arrestins / Q000235:genetics / Q000378:metabolism*; D001692:Biological Transport; D002460:Cell Line; D004705:Endocytosis; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D051381:Rats; D018040:Receptors, Neurokinin-1 / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013373:Substance P / Q000378:metabolism*; D014162:Transfection; D000071557:beta-Arrestins
D019390:Arrestins; D008164:Luminescent Proteins; D018040:Receptors, Neurokinin-1; D011993:Recombinant Fusion Proteins; D000071557:beta-Arrestins; D049452:Green Fluorescent Proteins; D013373:Substance P