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PMID: 11080148 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Dual interaction of synaptotagmin with mu2- and alpha-adaptin facilitates clathrin-coated pit nucleation.

The EMBO journal ·Vol. 19 ·No. 22 ·2000-11-15 ·Pages 6011-9

Haucke V, Wenk MR, Chapman ER, Farsad K, De Camilli P

Abstract

The synaptic vesicle protein synaptotagmin was proposed to act as a major docking site for the recruitment of clathrin coats implicated in endocytosis, including the recycling of synaptic vesicles. We show here that the C2B domain of synaptotagmin binds mu2- and alpha-adaptin, two of the four subunits of the endocytic adaptor complex AP-2. mu2 represents the major interacting subunit of AP-2 within this complex. Its binding to synaptotagmin is mediated by a site in subdomain B that is distinct from the binding site for tyrosine-based sorting motifs located in subdomain A. The presence of the C2B domain of synaptotagmin at the surface of liposomes enhances the recruitment of AP-2 and clathrin. Conversely, perturbation of the interaction between synaptotagmin and AP-2 by synprint, the cytoplasmic synaptotagmin-binding domain of N-type calcium channels, inhibits transferrin internalization in living cells. We conclude that a dual interaction of synaptotagmin with the clathrin adaptor AP-2 plays a key physiological role in the nucleation of endocytic clathrin-coated pits.

MeSH Terms
Adaptor Protein Complex 1 Adaptor Protein Complex 2 Adaptor Protein Complex 3 Adaptor Protein Complex alpha Subunits Adaptor Protein Complex mu Subunits Adaptor Proteins, Vesicular Transport Animals Binding Sites CHO Cells Calcium-Binding Proteins Clathrin/chemistry,metabolism Coated Pits, Cell-Membrane/metabolism Cricetinae In Vitro Techniques Liposomes Lysine/chemistry Membrane Glycoproteins/chemistry,metabolism Membrane Proteins/chemistry,genetics,metabolism Mutation Nerve Tissue Proteins/chemistry,genetics,metabolism Phosphoproteins/chemistry,genetics,metabolism Protein Subunits Rats Synaptic Vesicles/metabolism Synaptotagmins Tyrosine/chemistry
Chemicals
Adaptor Protein Complex 1 Adaptor Protein Complex 2 Adaptor Protein Complex 3 Adaptor Protein Complex alpha Subunits Adaptor Protein Complex mu Subunits Adaptor Proteins, Vesicular Transport Ap3m2 protein, rat Calcium-Binding Proteins Clathrin Liposomes Membrane Glycoproteins Membrane Proteins Nerve Tissue Proteins Phosphoproteins Protein Subunits adaptor protein complex 1, mu 2 subunit adaptor protein complex 2, mu 2 subunit Synaptotagmins Tyrosine Lysine
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Haucke V
Department of Cell Biology and Howard Hughes Medical Institute, Yale University School of Medicine, 295 Congress Avenue, New Haven, CT 06510, USA.
Wenk M R
Chapman E R
Farsad K
De Camilli P
References (48)
48 references, click to expand
  1. Interaction of the synprint site of N-type Ca2+ channels with the C2B domain of synaptotagmin I.
    Proc Natl Acad Sci U S A. 1997 May 13;94(10):5405-10 PMID: 9144250
  2. Regulatory interactions in the recognition of endocytic sorting signals by AP-2 complexes.
    EMBO J. 1997 May 1;16(9):2240-50 PMID: 9171339
  3. Synaptic vesicle endocytosis.
    Curr Opin Neurobiol. 1997 Jun;7(3):323-30 PMID: 9232811
  4. Linking cargo to vesicle formation: receptor tail interactions with coat proteins.
    Curr Opin Cell Biol. 1997 Aug;9(4):488-95 PMID: 9261055
  5. The role of ADP-ribosylation factor and phospholipase D in adaptor recruitment.
    J Cell Biol. 1997 Sep 22;138(6):1239-54 PMID: 9298980
  6. Ca2+-dependent and -independent interactions of the isoforms of the alpha1A subunit of brain Ca2+ channels with presynaptic SNARE proteins.
    Proc Natl Acad Sci U S A. 1997 Dec 23;94(26):14782-6 PMID: 9405690
  7. Generation of coated intermediates of clathrin-mediated endocytosis on protein-free liposomes.
    Cell. 1998 Jul 10;94(1):131-41 PMID: 9674434
  8. Clathrin and adaptors.
    Biochim Biophys Acta. 1998 Aug 14;1404(1-2):173-93 PMID: 9714795
  9. Dissociation between Ca2+-triggered synaptic vesicle exocytosis and clathrin-mediated endocytosis at a central synapse.
    Neuron. 1998 Sep;21(3):607-16 PMID: 9768846
  10. A structural explanation for the recognition of tyrosine-based endocytotic signals.
    Science. 1998 Nov 13;282(5392):1327-32 PMID: 9812899
  11. Delineation of the oligomerization, AP-2 binding, and synprint binding region of the C2B domain of synaptotagmin.
    J Biol Chem. 1998 Dec 4;273(49):32966-72 PMID: 9830048
  12. Coat assembly directs v-SNARE concentration into synthetic COPII vesicles.
    Mol Cell. 1998 Nov;2(5):703-8 PMID: 9844642
  13. Coupling of coat assembly and vesicle budding to packaging of putative cargo receptors.
    Cell. 1999 Feb 19;96(4):495-506 PMID: 10052452
  14. Impairment of dynamin's GAP domain stimulates receptor-mediated endocytosis.
    Nature. 1999 Apr 8;398(6727):481-6 PMID: 10206643
  15. A primer on vesicle budding.
    Cell. 1999 Apr 16;97(2):145-8 PMID: 10219233
  16. Inhibition of the receptor-binding function of clathrin adaptor protein AP-2 by dominant-negative mutant mu2 subunit and its effects on endocytosis.
    EMBO J. 1999 May 4;18(9):2489-99 PMID: 10228163
  17. Molecular bases for the recognition of tyrosine-based sorting signals.
    J Cell Biol. 1999 May 31;145(5):923-6 PMID: 10352010
  18. Coupled inositide phosphorylation and phospholipase D activation initiates clathrin-coat assembly on lysosomes.
    J Biol Chem. 1999 Jun 18;274(25):17794-805 PMID: 10364223
  19. The subcellular localizations of atypical synaptotagmins III and VI. Synaptotagmin III is enriched in synapses and synaptic plasma membranes but not in synaptic vesicles.
    J Biol Chem. 1999 Jun 25;274(26):18290-6 PMID: 10373432
  20. The structural era of endocytosis.
    Science. 1999 Jul 9;285(5425):215-20 PMID: 10398591
  21. AP-2 recruitment to synaptotagmin stimulated by tyrosine-based endocytic motifs.
    Science. 1999 Aug 20;285(5431):1268-71 PMID: 10455054
  22. Phosphoinositide-AP-2 interactions required for targeting to plasma membrane clathrin-coated pits.
    J Cell Biol. 1999 Aug 23;146(4):755-64 PMID: 10459011
  23. Endophilin I mediates synaptic vesicle formation by transfer of arachidonate to lysophosphatidic acid.
    Nature. 1999 Sep 9;401(6749):133-41 PMID: 10490020
  24. Essential role of phosphoinositide metabolism in synaptic vesicle recycling.
    Cell. 1999 Oct 15;99(2):179-88 PMID: 10535736
  25. Functional partnership between amphiphysin and dynamin in clathrin-mediated endocytosis.
    Nat Cell Biol. 1999 May;1(1):33-9 PMID: 10559861
  26. The endocytic machinery in nerve terminals surrounds sites of exocytosis.
    Curr Biol. 1999 Dec 2;9(23):1411-4 PMID: 10607569
  27. Endophilin/SH3p4 is required for the transition from early to late stages in clathrin-mediated synaptic vesicle endocytosis.
    Neuron. 1999 Sep;24(1):143-54 PMID: 10677033
  28. Synaptotagmin VII regulates Ca(2+)-dependent exocytosis of lysosomes in fibroblasts.
    J Cell Biol. 2000 Mar 20;148(6):1141-49 PMID: 10725327
  29. SCAMP1 function in endocytosis.
    J Biol Chem. 2000 Apr 28;275(17):12752-6 PMID: 10777571
  30. Synaptotagmin regulation of coated pit assembly.
    J Biol Chem. 2000 Oct 6;275(40):30916-24 PMID: 10906143
  31. Evidence for recycling of synaptic vesicle membrane during transmitter release at the frog neuromuscular junction.
    J Cell Biol. 1973 May;57(2):315-44 PMID: 4348786
  32. Effects of cytoplasmic acidification on clathrin lattice morphology.
    J Cell Biol. 1989 Feb;108(2):401-11 PMID: 2563729
  33. Clathrin-coated pits contain an integral membrane protein that binds the AP-2 subunit with high affinity.
    J Biol Chem. 1990 Sep 25;265(27):16514-20 PMID: 1975814
  34. Interaction of assembly protein AP-2 and its isolated subunits with clathrin.
    Biochemistry. 1991 Jun 4;30(22):5590-7 PMID: 2036430
  35. Clathrin assembly protein AP180: primary structure, domain organization and identification of a clathrin binding site.
    EMBO J. 1993 Feb;12(2):667-75 PMID: 8440257
  36. Synaptotagmin I is a high affinity receptor for clathrin AP-2: implications for membrane recycling.
    Cell. 1994 Sep 9;78(5):751-60 PMID: 8087843
  37. Tubular membrane invaginations coated by dynamin rings are induced by GTP-gamma S in nerve terminals.
    Nature. 1995 Mar 9;374(6518):186-90 PMID: 7877693
  38. Dynamin self-assembles into rings suggesting a mechanism for coated vesicle budding.
    Nature. 1995 Mar 9;374(6518):190-2 PMID: 7877694
  39. Association of epidermal growth factor receptors with coated pit adaptins via a tyrosine phosphorylation-regulated mechanism.
    J Biol Chem. 1995 Mar 17;270(11):6320-7 PMID: 7534311
  40. Clathrin binding and assembly activities of expressed domains of the synapse-specific clathrin assembly protein AP-3.
    J Biol Chem. 1995 May 5;270(18):10933-9 PMID: 7738035
  41. Ca(2+)-dependent and -independent activities of neural and non-neural synaptotagmins.
    Nature. 1995 Jun 15;375(6532):594-9 PMID: 7791877
  42. Interaction of tyrosine-based sorting signals with clathrin-associated proteins.
    Science. 1995 Sep 29;269(5232):1872-5 PMID: 7569928
  43. Role of the C2B domain of synaptotagmin in vesicular release and recycling as determined by specific antibody injection into the squid giant synapse preterminal.
    Proc Natl Acad Sci U S A. 1995 Nov 7;92(23):10708-12 PMID: 7479869
  44. Defective recycling of synaptic vesicles in synaptotagmin mutants of Caenorhabditis elegans.
    Nature. 1995 Nov 9;378(6553):196-9 PMID: 7477324
  45. A novel class of clathrin-coated vesicles budding from endosomes.
    J Cell Biol. 1996 Jan;132(1-2):21-33 PMID: 8567724
  46. Bimodal interaction of coatomer with the p24 family of putative cargo receptors.
    Science. 1996 Sep 6;273(5280):1396-9 PMID: 8703076
  47. Endocytosis and molecular sorting.
    Annu Rev Cell Dev Biol. 1996;12:575-625 PMID: 8970738
  48. Synaptic vesicle endocytosis impaired by disruption of dynamin-SH3 domain interactions.
    Science. 1997 Apr 11;276(5310):259-63 PMID: 9092476
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/19.22.6011
Published
2000-11-15
Pages
6011-9
Language
English
Region
England
NLM ID
8208664
PMCID
PMC305843
Subset
IM
Grants
NIGMS NIH HHS · R01 GM056827 · United States
NINDS NIH HHS · NS33709 · United States
NINDS NIH HHS · NS36252 · United States
NIGMS NIH HHS · R01GM56827-01 · United States
External Links
PubMed source DOI: 10.1093/emboj/19.22.6011 PMC Full Text View journal details
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