The temporal profile of activity-dependent presynaptic phospho-signalling reveals long-lasting patterns of poststimulus regulation

Engholm-Keller, Kasper, Waardenberg, Ashley J., Müller, Johannes A., Wark, Jesse R., Fernando, Rowena N., Arthur, Jonathan W., Robinson, Phillip J., Dietrich, Dirk, Schoch, Susanne, Graham, Mark E., UNSPECIFIED, and UNSPECIFIED (2019) The temporal profile of activity-dependent presynaptic phospho-signalling reveals long-lasting patterns of poststimulus regulation. PLoS Biology, 17 (3). e3000170. (In Press)

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Abstract

Depolarization of presynaptic terminals stimulates calcium influx, which evokes neurotransmitter release and activates phosphorylation-based signalling. Here, we present the first global temporal profile of presynaptic activity-dependent phospho-signalling, which includes two KCl stimulation levels and analysis of the poststimulus period. We profiled 1,917 regulated phosphopeptides and bioinformatically identified six temporal patterns of co-regulated proteins. The presynaptic proteins with large changes in phospho-status were again prominently regulated in the analysis of 7,070 activity-dependent phosphopeptides from KCl-stimulated cultured hippocampal neurons. Active zone scaffold proteins showed a high level of activity-dependent phospho-regulation that far exceeded the response from postsynaptic density scaffold proteins. Accordingly, bassoon was identified as the major target of neuronal phospho-signalling. We developed a probabilistic computational method, KinSwing, which matched protein kinase substrate motifs to regulated phosphorylation sites to reveal underlying protein kinase activity. This approach allowed us to link protein kinases to profiles of co-regulated presynaptic protein networks. Ca2+- and calmodulin-dependent protein kinase IIα (CaMKIIα) responded rapidly, scaled with stimulus strength, and had long-lasting activity. Mitogen-activated protein kinase (MAPK)/extracellular signal–regulated kinase (ERK) was the main protein kinase predicted to control a distinct and significant pattern of poststimulus up-regulation of phosphorylation. This work provides a unique resource of activity-dependent phosphorylation sites of synaptosomes and neurons, the vast majority of which have not been investigated with regard to their functional impact. This resource will enable detailed characterization of the phospho-regulated mechanisms impacting the plasticity of neurotransmitter release.

Item ID: 57389
Item Type: Article (Research - C1)
ISSN: 1545-7885
Keywords: neuroscience, synaptic plasticity, kinase activity prediction, kinswing, phosphoproteomics
Copyright Information: Copyright © 2019 Engholm-Keller et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funders: Lundbeck Foundation (LF), Danish Council for Independent Research (DCIR), FP7 Marie Curie Actions – COFUND, Carlsberg Foundation (CF), National Health and Medical Research Council (NHMRC), Brain Foundation (BF), Australian Cancer Research Foundation (ACRF), Cancer Institute New South Wales, Zero Childhood Cancer, Ramaciotti Foundation (RF), Honda Foundation, Bruce Wall Estate, German Research Council (GRC), BONFOR, Rebecca L Cooper Medical Research Foundation
Projects and Grants: LF R83-2011-8143, FP7 Marie Curie Actions – COFUND DFF – 1325-00154, CF CF15-1056, CF CF16-0066, NHMRC 1079160, NHMRC 1077989, NHMRC 1052494, NHMRC 571070, GRC DFG, GRC SFB1089, GRC SPP1757, GRC SCHO 820/6-1, GRC SCHO 820/4-1, GRC DI853/3-2, GRC DI853/7-1
Date Deposited: 12 Mar 2019 02:27
FoR Codes: 11 MEDICAL AND HEALTH SCIENCES > 1109 Neurosciences > 110999 Neurosciences not elsewhere classified @ 35%
06 BIOLOGICAL SCIENCES > 0601 Biochemistry and Cell Biology > 060102 Bioinformatics @ 30%
06 BIOLOGICAL SCIENCES > 0601 Biochemistry and Cell Biology > 060109 Proteomics and Intermolecular Interactions (excl Medical Proteomics) @ 35%
SEO Codes: 97 EXPANDING KNOWLEDGE > 970106 Expanding Knowledge in the Biological Sciences @ 100%
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