Phosphorylation of synaptic vesicle protein 2A at Thr84 by casein kinase 1 family kinases controls the specific retrieval of synaptotagmin-1

Ning Zhang, Sarah L. Gordon, Maximilian J. Fritsch, Noor Esoof, David G. Campbell, Robert Gourlay, Velupillai Srikannathasan, Thomas Macartney, Mark Peggie, Daan M. F. van Aalten, Michael A. Cousin (Lead / Corresponding author), Dario R. Alessi (Lead / Corresponding author)

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    Abstract

    Synaptic vesicle protein 2A (SV2A) is a ubiquitous component of synaptic vesicles (SVs). It has roles in both SV trafficking and neurotransmitter release. We demonstrate that Casein kinase 1 family members, including isoforms of Tau–tubulin protein kinases (TTBK1 and TTBK2), phosphorylate human SV2A at two constellations of residues, namely Cluster-1 (Ser42, Ser45, and Ser47) and Cluster-2 (Ser80, Ser81, and Thr84). These residues are also phosphorylated in vivo, and the phosphorylation of Thr84 within Cluster-2 is essential for triggering binding to the C2B domain of human synaptotagmin-1. We show by crystallographic and other analyses that the phosphorylated Thr84 residue binds to a pocket formed by three conserved Lys residues (Lys314, Lys326, and Lys328) on the surface of the synaptotagmin-1 C2B domain. Finally, we observed dysfunctional synaptotagmin-1 retrieval during SV endocytosis by ablating its phospho-dependent interaction with SV2A, knockdown of SV2A, or rescue with a phosphorylation-null Thr84 SV2A mutant in primary cultures of mouse neurons. This study reveals fundamental details of how phosphorylation of Thr84 on SV2A controls its interaction with synaptotagmin-1 and implicates SV2A as a phospho-dependent chaperone required for the specific retrieval of synaptotagmin-1 during SV endocytosis.

    Original languageEnglish
    Pages (from-to)2492-2507
    Number of pages16
    JournalJournal of Neuroscience
    Volume35
    Issue number6
    DOIs
    Publication statusPublished - 11 Feb 2015

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    Keywords

    • CK1
    • SV2A
    • Synaptotagmin

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