5-HT2A receptor signalling through phospholipase D1 associated with its C-terminal tail

Zoe Barclay, Louise Dickson, Derek N. Robertson, Melanie S. Johnson, Pamela J. Holland, Roberta Rosie, Liting Sun, Susan Fleetwood-Walker, Eve M. Lutz, Rory Mitchell

Research output: Contribution to journalArticlepeer-review

Abstract

The 5-HT2AR (5-hydroxytryptamine-2A receptor) is a GPCR (G-protein-coupled receptor) that is implicated in the actions of hallucinogens and represents a major target of atypical antipsychotic agents. In addition to its classical signalling though PLC (phospholipase C), the receptor can activate several other pathways, including ARF (A DP-ribosylation factor)-dependent activation of PLD (phospholipase D), which appears to be achieved through a mechanism independent of heterotrimeric G-proteins. In the present study we show that wild-type and inactive constructs of PLD1 (but not PLD2) respectively facilitate and inhibit ARF-dependent PLD signalling by the 5-HT2AR. Furthermore we demonstrate that PLD I specifically co-immunoprecipitates with the receptor and binds to a distal site in GST (glutathione transferase) fusion protein constructs of its C-terminal tail which is distinct from the ARF-interaction site, thereby suggesting the existence of a functional ARF-PLD signalling complex directly associated with this receptor. This reveals the spatial co-ordination of an important GPCR, transducer and effector into a physical complex that is likely to reinforce the impact of receptor activation on a heterotrimeric G-protein-independent signalling pathway. Signalling of this receptor through such non-canonical pathways may be important to its role in particular disorders.

Original languageEnglish
Pages (from-to)651-660
Number of pages10
JournalBiochemical Journal
Volume436
DOIs
Publication statusPublished - 15 Jun 2011

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