Proteomic Identification of an Endogenous Synaptic SUMOylome in the Developing Rat Brain.

Marie Pronot,Raphaël Forquet,Félicie Kieffer,Stéphane Martin,Lenka Schorova,Anne-Sophie Gay,Carole Gwizdek,Delphine Debayle,Gwénola Poupon

doi:10.3389/fnmol.2021.780535

Abstract

Synapses are highly specialized structures that interconnect neurons to form functional networks dedicated to neuronal communication. During brain development, synapses undergo activity-dependent rearrangements leading to both structural and functional changes. Many molecular processes are involved in this regulation, including post-translational modifications by the Small Ubiquitin-like MOdifier SUMO. To get a wider view of the panel of endogenous synaptic SUMO-modified proteins in the mammalian brain, we combined subcellular fractionation of rat brains at the post-natal day 14 with denaturing immunoprecipitation using SUMO2/3 antibodies and tandem mass spectrometry analysis. Our screening identified 803 candidate SUMO2/3 targets, which represents about 18% of the synaptic proteome. Our dataset includes neurotransmitter receptors, transporters, adhesion molecules, scaffolding proteins as well as vesicular trafficking and cytoskeleton-associated proteins, defining SUMO2/3 as a central regulator of the synaptic organization and function.

Highlights

A functional neuronal network relies on the coordinated organization of billions of highly specified contact points called synapses that interconnect neurons and allow communication in the mammalian brain
We identified 803 candidate SUMO2/3-modified proteins at the synapses of PND14 rat brains by a proteomic approach
Earlier studies using the double-tagged His6HA-SUMO1 knock-in mice concluded that SUMO1-ylated proteins are not present at synapses (Tirard et al, 2012; Daniel et al, 2017)

Summary

Introduction

A functional neuronal network relies on the coordinated organization of billions of highly specified contact points called synapses that interconnect neurons and allow communication in the mammalian brain. Synapses undergo constant activitydependent protein content rearrangements leading to both structural and functional changes. Many processes are involved in this modulation, including proteins, mRNAs and organelle transports along axons and dendrites as well as synaptic vesicular trafficking, synaptodendritic exchanges, local translation and protein degradation. Post-translational modifications (PTM) such as SUMOylation regulate protein function as well as the shaping of macromolecular complexes and participate in those processes. SUMO conjugation is achieved by the sole E2 SUMO-conjugating enzyme Ubc. Substrate recognition by Ubc is either direct or mediated by bridging proteins called E3 ligases. There are three SUMO paralogs (SUMO 1, 2, and 3) in the mammalian brain.

Methods

Results

Conclusion