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

Zoe Barclay , Louise Dickson , Derek Robertson , Melanie Johnson , Pamela Holland , Roberta Rosie , Liting Sun , Sue Fleetwood-Walker , Eve M Lutz , Rory Mitchell (1)

1 Centre for Integrative Physiology

The 5-hydroxytryptamine-2A receptor (5-HT2AR) is a G protein-coupled receptor (GPCR) that is implicated in the actions of hallucinogens and represents a major target of atypical antipsychotic agents. In addition to its classical signalling though phospholipase C (PLC), the receptor can activate several other pathways, including ARF-dependent activation of phospholipase D (PLD), which appears to be achieved through a mechanism independent of heterotrimeric G proteins. We show here that wild type and inactive constructs of PLD1 (but not PLD2) respectively facilitate and inhibit ARF-dependent PLD signalling by the 5-HT2AR. Further we demonstrate that PLD1 specifically co-immunoprecipitates with the receptor and binds to a distal site in GST-fusion protein constructs of its carboxy-terminal tail that 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 particular heterotrimeric G protein-independent signaling pathway. Signalling of this receptor through such non-canonical pathways may be important to its role in particular disorders.

Mots clés: Life Sciences

Zoe Barclay, Louise Dickson, Derek Robertson, Melanie Johnson, Pamela Holland, et al.. 5-HT2A receptor signalling through phospholipase D1 associated with its carboxy-terminal tail. Biochemical Journal, 2011, 436 (3), pp.651-660. ⟨10.1042/BJ20101844⟩. ⟨hal-00596269⟩