Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1996 Jan 9;93(1):331–335. doi: 10.1073/pnas.93.1.331

A role of amphiphysin in synaptic vesicle endocytosis suggested by its binding to dynamin in nerve terminals.

C David 1, P S McPherson 1, O Mundigl 1, P de Camilli 1
PMCID: PMC40232  PMID: 8552632

Abstract

Amphiphysin, a major autoantigen in paraneoplastic Stiff-Man syndrome, is an SH3 domain-containing neuronal protein, concentrated in nerve terminals. Here, we demonstrate a specific, SH3 domain-mediated, interaction between amphiphysin and dynamin by gel overlay and affinity chromatography. In addition, we show that the two proteins are colocalized in nerve terminals and are coprecipitated from brain extracts consistent with their interactions in situ. We also report that a region of amphiphysin distinct from its SH3 domain mediates its binding to the alpha c subunit of AP2 adaptin, which is also concentrated in nerve terminals. These findings support a role of amphiphysin in synaptic vesicle endocytosis.

Full text

PDF
331

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amberg D. C., Basart E., Botstein D. Defining protein interactions with yeast actin in vivo. Nat Struct Biol. 1995 Jan;2(1):28–35. doi: 10.1038/nsb0195-28. [DOI] [PubMed] [Google Scholar]
  2. Ball C. L., Hunt S. P., Robinson M. S. Expression and localization of alpha-adaptin isoforms. J Cell Sci. 1995 Aug;108(Pt 8):2865–2875. doi: 10.1242/jcs.108.8.2865. [DOI] [PubMed] [Google Scholar]
  3. Bauer F., Urdaci M., Aigle M., Crouzet M. Alteration of a yeast SH3 protein leads to conditional viability with defects in cytoskeletal and budding patterns. Mol Cell Biol. 1993 Aug;13(8):5070–5084. doi: 10.1128/mcb.13.8.5070. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chen M. S., Obar R. A., Schroeder C. C., Austin T. W., Poodry C. A., Wadsworth S. C., Vallee R. B. Multiple forms of dynamin are encoded by shibire, a Drosophila gene involved in endocytosis. Nature. 1991 Jun 13;351(6327):583–586. doi: 10.1038/351583a0. [DOI] [PubMed] [Google Scholar]
  5. Chin D. J., Straubinger R. M., Acton S., Näthke I., Brodsky F. M. 100-kDa polypeptides in peripheral clathrin-coated vesicles are required for receptor-mediated endocytosis. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9289–9293. doi: 10.1073/pnas.86.23.9289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Crouzet M., Urdaci M., Dulau L., Aigle M. Yeast mutant affected for viability upon nutrient starvation: characterization and cloning of the RVS161 gene. Yeast. 1991 Oct;7(7):727–743. doi: 10.1002/yea.320070708. [DOI] [PubMed] [Google Scholar]
  7. Damke H., Baba T., Warnock D. E., Schmid S. L. Induction of mutant dynamin specifically blocks endocytic coated vesicle formation. J Cell Biol. 1994 Nov;127(4):915–934. doi: 10.1083/jcb.127.4.915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. David C., Solimena M., De Camilli P. Autoimmunity in stiff-Man syndrome with breast cancer is targeted to the C-terminal region of human amphiphysin, a protein similar to the yeast proteins, Rvs167 and Rvs161. FEBS Lett. 1994 Aug 29;351(1):73–79. doi: 10.1016/0014-5793(94)00826-4. [DOI] [PubMed] [Google Scholar]
  9. De Camilli P., Harris S. M., Jr, Huttner W. B., Greengard P. Synapsin I (Protein I), a nerve terminal-specific phosphoprotein. II. Its specific association with synaptic vesicles demonstrated by immunocytochemistry in agarose-embedded synaptosomes. J Cell Biol. 1983 May;96(5):1355–1373. doi: 10.1083/jcb.96.5.1355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. De Camilli P., Takei K., McPherson P. S. The function of dynamin in endocytosis. Curr Opin Neurobiol. 1995 Oct;5(5):559–565. doi: 10.1016/0959-4388(95)80059-x. [DOI] [PubMed] [Google Scholar]
  11. De Camilli P., Thomas A., Cofiell R., Folli F., Lichte B., Piccolo G., Meinck H. M., Austoni M., Fassetta G., Bottazzo G. The synaptic vesicle-associated protein amphiphysin is the 128-kD autoantigen of Stiff-Man syndrome with breast cancer. J Exp Med. 1993 Dec 1;178(6):2219–2223. doi: 10.1084/jem.178.6.2219. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Desfarges L., Durrens P., Juguelin H., Cassagne C., Bonneu M., Aigle M. Yeast mutants affected in viability upon starvation have a modified phospholipid composition. Yeast. 1993 Mar;9(3):267–277. doi: 10.1002/yea.320090306. [DOI] [PubMed] [Google Scholar]
  13. Folli F., Solimena M., Cofiell R., Austoni M., Tallini G., Fassetta G., Bates D., Cartlidge N., Bottazzo G. F., Piccolo G. Autoantibodies to a 128-kd synaptic protein in three women with the stiff-man syndrome and breast cancer. N Engl J Med. 1993 Feb 25;328(8):546–551. doi: 10.1056/NEJM199302253280805. [DOI] [PubMed] [Google Scholar]
  14. Garcia E. P., Gatti E., Butler M., Burton J., De Camilli P. A rat brain Sec1 homologue related to Rop and UNC18 interacts with syntaxin. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2003–2007. doi: 10.1073/pnas.91.6.2003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gout I., Dhand R., Hiles I. D., Fry M. J., Panayotou G., Das P., Truong O., Totty N. F., Hsuan J., Booker G. W. The GTPase dynamin binds to and is activated by a subset of SH3 domains. Cell. 1993 Oct 8;75(1):25–36. [PubMed] [Google Scholar]
  16. Herskovits J. S., Burgess C. C., Obar R. A., Vallee R. B. Effects of mutant rat dynamin on endocytosis. J Cell Biol. 1993 Aug;122(3):565–578. doi: 10.1083/jcb.122.3.565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Herskovits J. S., Shpetner H. S., Burgess C. C., Vallee R. B. Microtubules and Src homology 3 domains stimulate the dynamin GTPase via its C-terminal domain. Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11468–11472. doi: 10.1073/pnas.90.24.11468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hinshaw J. E., Schmid S. L. Dynamin self-assembles into rings suggesting a mechanism for coated vesicle budding. Nature. 1995 Mar 9;374(6518):190–192. doi: 10.1038/374190a0. [DOI] [PubMed] [Google Scholar]
  19. Koenig J. H., Ikeda K. Disappearance and reformation of synaptic vesicle membrane upon transmitter release observed under reversible blockage of membrane retrieval. J Neurosci. 1989 Nov;9(11):3844–3860. doi: 10.1523/JNEUROSCI.09-11-03844.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kosaka T., Ikeda K. Possible temperature-dependent blockage of synaptic vesicle recycling induced by a single gene mutation in Drosophila. J Neurobiol. 1983 May;14(3):207–225. doi: 10.1002/neu.480140305. [DOI] [PubMed] [Google Scholar]
  21. Li C., Ullrich B., Zhang J. Z., Anderson R. G., Brose N., Südhof T. C. Ca(2+)-dependent and -independent activities of neural and non-neural synaptotagmins. Nature. 1995 Jun 15;375(6532):594–599. doi: 10.1038/375594a0. [DOI] [PubMed] [Google Scholar]
  22. Lichte B., Veh R. W., Meyer H. E., Kilimann M. W. Amphiphysin, a novel protein associated with synaptic vesicles. EMBO J. 1992 Jul;11(7):2521–2530. doi: 10.1002/j.1460-2075.1992.tb05317.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lowenstein E. J., Daly R. J., Batzer A. G., Li W., Margolis B., Lammers R., Ullrich A., Skolnik E. Y., Bar-Sagi D., Schlessinger J. The SH2 and SH3 domain-containing protein GRB2 links receptor tyrosine kinases to ras signaling. Cell. 1992 Aug 7;70(3):431–442. doi: 10.1016/0092-8674(92)90167-b. [DOI] [PubMed] [Google Scholar]
  24. McPherson P. S., Czernik A. J., Chilcote T. J., Onofri F., Benfenati F., Greengard P., Schlessinger J., De Camilli P. Interaction of Grb2 via its Src homology 3 domains with synaptic proteins including synapsin I. Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6486–6490. doi: 10.1073/pnas.91.14.6486. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. McPherson P. S., Takei K., Schmid S. L., De Camilli P. p145, a major Grb2-binding protein in brain, is co-localized with dynamin in nerve terminals where it undergoes activity-dependent dephosphorylation. J Biol Chem. 1994 Dec 2;269(48):30132–30139. [PubMed] [Google Scholar]
  26. Ohno H., Stewart J., Fournier M. C., Bosshart H., Rhee I., Miyatake S., Saito T., Gallusser A., Kirchhausen T., Bonifacino J. S. Interaction of tyrosine-based sorting signals with clathrin-associated proteins. Science. 1995 Sep 29;269(5232):1872–1875. doi: 10.1126/science.7569928. [DOI] [PubMed] [Google Scholar]
  27. Pawson T., Gish G. D. SH2 and SH3 domains: from structure to function. Cell. 1992 Oct 30;71(3):359–362. doi: 10.1016/0092-8674(92)90504-6. [DOI] [PubMed] [Google Scholar]
  28. Riezman H. Yeast endocytosis. Trends Cell Biol. 1993 Aug;3(8):273–277. doi: 10.1016/0962-8924(93)90056-7. [DOI] [PubMed] [Google Scholar]
  29. Robinson M. S. 100-kD coated vesicle proteins: molecular heterogeneity and intracellular distribution studied with monoclonal antibodies. J Cell Biol. 1987 Apr;104(4):887–895. doi: 10.1083/jcb.104.4.887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Robinson M. S. Cloning of cDNAs encoding two related 100-kD coated vesicle proteins (alpha-adaptins). J Cell Biol. 1989 Mar;108(3):833–842. doi: 10.1083/jcb.108.3.833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Robinson M. S. The role of clathrin, adaptors and dynamin in endocytosis. Curr Opin Cell Biol. 1994 Aug;6(4):538–544. doi: 10.1016/0955-0674(94)90074-4. [DOI] [PubMed] [Google Scholar]
  32. Rozakis-Adcock M., Fernley R., Wade J., Pawson T., Bowtell D. The SH2 and SH3 domains of mammalian Grb2 couple the EGF receptor to the Ras activator mSos1. Nature. 1993 May 6;363(6424):83–85. doi: 10.1038/363083a0. [DOI] [PubMed] [Google Scholar]
  33. Seedorf K., Kostka G., Lammers R., Bashkin P., Daly R., Burgess W. H., van der Bliek A. M., Schlessinger J., Ullrich A. Dynamin binds to SH3 domains of phospholipase C gamma and GRB-2. J Biol Chem. 1994 Jun 10;269(23):16009–16014. [PubMed] [Google Scholar]
  34. Shpetner H. S., Vallee R. B. Identification of dynamin, a novel mechanochemical enzyme that mediates interactions between microtubules. Cell. 1989 Nov 3;59(3):421–432. doi: 10.1016/0092-8674(89)90027-5. [DOI] [PubMed] [Google Scholar]
  35. Sivadon P., Bauer F., Aigle M., Crouzet M. Actin cytoskeleton and budding pattern are altered in the yeast rvs161 mutant: the Rvs161 protein shares common domains with the brain protein amphiphysin. Mol Gen Genet. 1995 Feb 20;246(4):485–495. doi: 10.1007/BF00290452. [DOI] [PubMed] [Google Scholar]
  36. Sorkin A., McKinsey T., Shih W., Kirchhausen T., Carpenter G. Stoichiometric interaction of the epidermal growth factor receptor with the clathrin-associated protein complex AP-2. J Biol Chem. 1995 Jan 13;270(2):619–625. doi: 10.1074/jbc.270.2.619. [DOI] [PubMed] [Google Scholar]
  37. Takei K., McPherson P. S., Schmid S. L., De Camilli P. Tubular membrane invaginations coated by dynamin rings are induced by GTP-gamma S in nerve terminals. Nature. 1995 Mar 9;374(6518):186–190. doi: 10.1038/374186a0. [DOI] [PubMed] [Google Scholar]
  38. Vallee R. B., Okamoto P. M. The regulation of endocytosis: identifying dynamin's binding partners. Trends Cell Biol. 1995 Feb;5(2):43–47. doi: 10.1016/s0962-8924(00)88937-0. [DOI] [PubMed] [Google Scholar]
  39. Wang L. H., Südhof T. C., Anderson R. G. The appendage domain of alpha-adaptin is a high affinity binding site for dynamin. J Biol Chem. 1995 Apr 28;270(17):10079–10083. doi: 10.1074/jbc.270.17.10079. [DOI] [PubMed] [Google Scholar]
  40. Zhang J. Z., Davletov B. A., Südhof T. C., Anderson R. G. Synaptotagmin I is a high affinity receptor for clathrin AP-2: implications for membrane recycling. Cell. 1994 Sep 9;78(5):751–760. doi: 10.1016/s0092-8674(94)90442-1. [DOI] [PubMed] [Google Scholar]
  41. van der Bliek A. M., Meyerowitz E. M. Dynamin-like protein encoded by the Drosophila shibire gene associated with vesicular traffic. Nature. 1991 May 30;351(6325):411–414. doi: 10.1038/351411a0. [DOI] [PubMed] [Google Scholar]
  42. van der Bliek A. M., Redelmeier T. E., Damke H., Tisdale E. J., Meyerowitz E. M., Schmid S. L. Mutations in human dynamin block an intermediate stage in coated vesicle formation. J Cell Biol. 1993 Aug;122(3):553–563. doi: 10.1083/jcb.122.3.553. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES