Abstract
Dynamin I is dephosphorylated at Ser-774 and Ser-778 during synaptic vesicle endocytosis (SVE) in nerve terminals. Phosphorylation was proposed to regulate the assembly of an endocytic protein complex with amphiphysin or endophilin. Instead, we found it recruits syndapin I for SVE and does not control amphiphysin or endophilin binding in rat synaptosomes. After depolarization, syndapin showed a calcineurin-mediated interaction with dynamin. A peptide mimicking the phosphorylation sites disrupted the dynamin-syndapin complex, not the dynamin-endophilin complex, arrested SVE and produced glutamate release fatigue after repetitive stimulation. Pseudophosphorylation of Ser-774 or Ser-778 inhibited syndapin binding without affecting amphiphysin recruitment. Site mutagenesis to alanine arrested SVE in cultured neurons. The effects of the sites were additive for syndapin I binding and SVE. Thus syndapin I is a central component of the endocytic protein complex for SVE via stimulus-dependent recruitment to dynamin I and has a key role in synaptic transmission.
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Acknowledgements
We thank the large number of our colleagues who have generously provided materials for this study. We particularly wish to thank J. Xue, A. Quan and G. Evans for technical advice and assistance, and E. van Dam, R. Duncan, M. Shipston and P. Rowe for critical reading of the manuscript. This work was supported by grants from the National Health and Medical Research Council of Australia, the Wellcome Trust (Ref: GR070569), an Australian Bicentennial Scholarship (V.A.), a University of Sydney Postgraduate Award (V.A.) and a University of Edinburgh Medical Faculty Scholarship (K.J.S.).
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Supplementary Fig. 1
DynIdmA and DynIdmE cause SVE arrest in cerebellar granule neurons. (PDF 109 kb)
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Anggono, V., Smillie, K., Graham, M. et al. Syndapin I is the phosphorylation-regulated dynamin I partner in synaptic vesicle endocytosis. Nat Neurosci 9, 752–760 (2006). https://doi.org/10.1038/nn1695
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DOI: https://doi.org/10.1038/nn1695
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