Differential modulation of collybistin conformational dynamics by the closely related GTPases Cdc42 and TC10

Nasir Imam; Susobhan Choudhury; Katrin G Heinze; Hermann Schindelin

doi:10.3389/fnsyn.2022.959875

Differential modulation of collybistin conformational dynamics by the closely related GTPases Cdc42 and TC10

Front Synaptic Neurosci. 2022 Aug 4:14:959875. doi: 10.3389/fnsyn.2022.959875. eCollection 2022.

Authors

Nasir Imam¹, Susobhan Choudhury², Katrin G Heinze², Hermann Schindelin¹

Affiliations

¹ Institute of Structural Biology, Rudolf Virchow Center for Integrative and Translational Bioimaging, University of Würzburg, Würzburg, Germany.
² Molecular Microscopy, Rudolf Virchow Center for Integrative and Translational Bioimaging, University of Würzburg, Würzburg, Germany.

Abstract

Interneuronal synaptic transmission relies on the proper spatial organization of presynaptic neurotransmitter release and its reception on the postsynaptic side by cognate neurotransmitter receptors. Neurotransmitter receptors are incorporated into and arranged within the plasma membrane with the assistance of scaffolding and adaptor proteins. At inhibitory GABAergic postsynapses, collybistin, a neuronal adaptor protein, recruits the scaffolding protein gephyrin and interacts with various neuronal factors including cell adhesion proteins of the neuroligin family, the GABA _A receptor α2-subunit and the closely related small GTPases Cdc42 and TC10 (RhoQ). Most collybistin splice variants harbor an N-terminal SH3 domain and exist in an autoinhibited/closed state. Cdc42 and TC10, despite sharing 67.4% amino acid sequence identity, interact differently with collybistin. Here, we delineate the molecular basis of the collybistin conformational activation induced by TC10 with the aid of recently developed collybistin FRET sensors. Time-resolved fluorescence-based FRET measurements reveal that TC10 binds to closed/inactive collybistin leading to relief of its autoinhibition, contrary to Cdc42, which only interacts with collybistin when forced into an open state by the introduction of mutations destabilizing the closed state of collybistin. Taken together, our data describe a TC10-driven signaling mechanism in which collybistin switches from its autoinhibited closed state to an open/active state.

Keywords: Rho GTPase; autoinhibition; fluorescence resonance energy transfer (FRET); gephyrin; guanine nucleotide exchange factor (GEF); inhibitory postsynapse.