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)

    Research output: Contribution to journalArticle

    28 Citations (Scopus)

    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

    Fingerprint

    Synaptotagmin I
    Casein Kinase I
    Synaptic Vesicles
    Phosphotransferases
    Phosphorylation
    Proteins
    Endocytosis
    Mutant Proteins
    Protein Kinases
    Neurotransmitter Agents
    Protein Isoforms

    Keywords

    • CK1
    • SV2A
    • Synaptotagmin

    Cite this

    Zhang, Ning ; Gordon, Sarah L. ; Fritsch, Maximilian J. ; Esoof, Noor ; Campbell, David G. ; Gourlay, Robert ; Srikannathasan, Velupillai ; Macartney, Thomas ; Peggie, Mark ; van Aalten, Daan M. F. ; Cousin, Michael A. ; Alessi, Dario R. / Phosphorylation of synaptic vesicle protein 2A at Thr84 by casein kinase 1 family kinases controls the specific retrieval of synaptotagmin-1. In: Journal of Neuroscience. 2015 ; Vol. 35, No. 6. pp. 2492-2507.
    @article{b3ba46c3374c48fabd723b5b80ad3333,
    title = "Phosphorylation of synaptic vesicle protein 2A at Thr84 by casein kinase 1 family kinases controls the specific retrieval of synaptotagmin-1",
    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.",
    keywords = "CK1, SV2A, Synaptotagmin",
    author = "Ning Zhang and Gordon, {Sarah L.} and Fritsch, {Maximilian J.} and Noor Esoof and Campbell, {David G.} and Robert Gourlay and Velupillai Srikannathasan and Thomas Macartney and Mark Peggie and {van Aalten}, {Daan M. F.} and Cousin, {Michael A.} and Alessi, {Dario R.}",
    year = "2015",
    month = "2",
    day = "11",
    doi = "10.1523/JNEUROSCI.4248-14.2015",
    language = "English",
    volume = "35",
    pages = "2492--2507",
    journal = "Journal of Neuroscience",
    issn = "0270-6474",
    publisher = "Society for Neuroscience",
    number = "6",

    }

    Phosphorylation of synaptic vesicle protein 2A at Thr84 by casein kinase 1 family kinases controls the specific retrieval of synaptotagmin-1. / Zhang, Ning; Gordon, Sarah L.; Fritsch, Maximilian J.; Esoof, Noor; Campbell, David G.; Gourlay, Robert; Srikannathasan, Velupillai; Macartney, Thomas; Peggie, Mark; van Aalten, Daan M. F.; Cousin, Michael A. (Lead / Corresponding author); Alessi, Dario R. (Lead / Corresponding author).

    In: Journal of Neuroscience, Vol. 35, No. 6, 11.02.2015, p. 2492-2507.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Phosphorylation of synaptic vesicle protein 2A at Thr84 by casein kinase 1 family kinases controls the specific retrieval of synaptotagmin-1

    AU - Zhang, Ning

    AU - Gordon, Sarah L.

    AU - Fritsch, Maximilian J.

    AU - Esoof, Noor

    AU - Campbell, David G.

    AU - Gourlay, Robert

    AU - Srikannathasan, Velupillai

    AU - Macartney, Thomas

    AU - Peggie, Mark

    AU - van Aalten, Daan M. F.

    AU - Cousin, Michael A.

    AU - Alessi, Dario R.

    PY - 2015/2/11

    Y1 - 2015/2/11

    N2 - 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.

    AB - 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.

    KW - CK1

    KW - SV2A

    KW - Synaptotagmin

    UR - http://www.scopus.com/inward/record.url?scp=84922567981&partnerID=8YFLogxK

    U2 - 10.1523/JNEUROSCI.4248-14.2015

    DO - 10.1523/JNEUROSCI.4248-14.2015

    M3 - Article

    C2 - 25673844

    AN - SCOPUS:84922567981

    VL - 35

    SP - 2492

    EP - 2507

    JO - Journal of Neuroscience

    JF - Journal of Neuroscience

    SN - 0270-6474

    IS - 6

    ER -