Multisite phosphorylation is required for sustained interaction with GRKs and arrestins during rapid -opioid receptor desensitization

Miess, Elke; Gondin, Arisbel B.; Yousuf, Arsalan; Steinborn, Ralph; M�sslein, Nadja; Yang, Yunshi; G�ldner, Martin; Ruland, Julia G.; B�nemann, Moritz; Krasel, Cornelius; Christie, MacDonald J.; Halls, Michelle L.; Schulz, Stefan; Canals, Meritxell

Abstract

Copyright © 2018 The Authors. G protein receptor kinases (GRKs) and -arrestins are key regulators of -opioid receptor (MOR) signaling and trafficking. We have previously shown that high-efficacy opioids such as DAMGO stimulate a GRK2/3-mediated multisite phosphorylation of conserved C-terminal tail serine and threonine residues, which facilitates internalization of the receptor. In contrast, morphine-induced phosphorylation of MOR is limited to Ser375 and is not sufficient to drive substantial receptor internalization. We report how specific multisite phosphorylation controlled the dynamics of GRK and -arrestin interactions with MOR and show how such phosphorylation mediated receptor desensitization. We showed that GRK2/3 was recruited more quickly than was -arrestin to a DAMGO-activated MOR. -Arrestin recruitment required GRK2 activity and MOR phosphorylation, but GRK recruitment also depended on the phosphorylation sites in the C-terminal tail, specifically four serine and threonine residues within the 370TREHPSTANT379 motif. Our results also suggested that other residues outside this motif participated in the initial and transient recruitment of GRK and -arrestins. We identified two components of high-efficacy agonist desensitization of MOR: a sustained component, which required GRK2-mediated phosphorylation and a potential soluble factor, and a rapid component, which was likely mediated by GRK2 but independent of receptor phosphorylation. Elucidating these complex receptor-effector interactions represents an important step toward a mechanistic understanding of MOR desensitization that leads to the development of tolerance and dependence.

Citation

Miess, E., Gondin, A. B., Yousuf, A., Steinborn, R., Mösslein, N., Yang, Y., Göldner, M., Ruland, J. G., Bünemann, M., Krasel, C., Christie, M. J., Halls, M. L., Schulz, S., & Canals, M. (2018). Multisite phosphorylation is required for sustained interaction with GRKs and arrestins during rapid -opioid receptor desensitization. Science Signaling, 11(539), Article eaas9609. https://doi.org/10.1126/scisignal.aas9609

• Journal Article Type: Article
• Acceptance Date: Jun 29, 2018
• Online Publication Date: Jul 17, 2018
• Publication Date: Jul 17, 2018
• Deposit Date: Mar 6, 2019
• Publicly Available Date: Mar 6, 2019
• Journal: Science Signaling
• Print ISSN: 1945-0877
• Electronic ISSN: 1937-9145
• Publisher: American Association for the Advancement of Science
• Peer Reviewed: Peer Reviewed
• Volume: 11
• Issue: 539
• Article Number: eaas9609
• DOI: https://doi.org/10.1126/scisignal.aas9609
• Public URL: https://nottingham-repository.worktribe.com/output/1311685
• Publisher URL: http://stke.sciencemag.org/content/11/539/eaas9609
• Related Public URLs: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85050106700&doi=10.1126%2fscisignal.aas9609&partnerID=40&md5=b697c108d3428d51248f991b199c04b9
• Additional Information: This is the author’s version of the work. It is posted here by permission of the AAAS for personal use, not for redistribution. The definitive version was published in Science Signaling on 17.07.2018; Vol. 11, issue 539, DOI: 10.1126/scisignal.aas9609
• Contract Date: Mar 6, 2019