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PMID: 1374414 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Specific association of the proto-oncogene product pp60c-src with an intracellular organelle, the PC12 synaptic vesicle.

The Journal of cell biology ·Vol. 117 ·No. 5 ·1992-06-00 ·Pages 1077-84

Linstedt AD, Vetter ML, Bishop JM, Kelly RB

Abstract

The protein product of the proto-oncogene c-src is a membrane-associated tyrosine kinase of unknown function. Identification of pp60c-src target membranes may elucidate the function of the c-src protein. The available evidence indicates that pp60c-src associates with distinct membranes within single cell types and has different distributions in different cell types. Our experiments demonstrate targeting of pp60c-src to an isolatable and biochemically identified membrane fraction in the neuroendocrine cell line PC12. The c-src protein was found to be specifically associated with synaptic vesicles since: (a) the pp60c-src immunofluorescent pattern overlapped with a synaptic vesicle marker, synaptophysin; (b) a significant proportion (44%) of the pp60c-src from PC12 but not fibroblast postnuclear supernatants was recovered in a small vesicle fraction; (c) an anti-synaptophysin cytoplasmic domain antibody immunodepleted all of the pp60c-src vesicles in this fraction, and (d) pp60c-src copurified during a 100-fold purification of PC12 synaptic vesicles. These results suggest a role for the c-src protein in the regulation of synaptic vesicle function.

MeSH Terms
Animals Fluorescent Antibody Technique Immunoblotting PC12 Cells Proto-Oncogene Proteins pp60(c-src)/analysis Synaptic Vesicles/chemistry Synaptophysin/analysis
Chemicals
Synaptophysin Proto-Oncogene Proteins pp60(c-src)
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Linstedt A D
Program in Neuroscience, University of California, San Francisco 94143-0534.
Vetter M L
Bishop J M
Kelly R B
References (40)
40 references, click to expand
  1. Polyomavirus middle T antigen downregulates junctional cell-to-cell communication.
    Mol Cell Biol. 1987 Feb;7(2):946-50 PMID: 2434836
  2. Isolation of monoclonal antibodies that recognize the transforming proteins of avian sarcoma viruses.
    J Virol. 1983 Nov;48(2):352-60 PMID: 6312092
  3. Protein tyrosine kinase activity and its endogenous substrates in rat brain: a subcellular and regional survey.
    J Neurochem. 1988 May;50(5):1447-55 PMID: 2452235
  4. c-src gene product in developing rat brain is enriched in nerve growth cone membranes.
    Proc Natl Acad Sci U S A. 1988 Jul;85(14):5001-5 PMID: 2455889
  5. p60c-src is complexed with a cellular protein in subcellular compartments involved in exocytosis.
    J Cell Biol. 1988 Dec;107(6 Pt 1):2125-35 PMID: 2461946
  6. Translocation of synapsin I in response to depolarization of isolated nerve terminals.
    Proc Natl Acad Sci U S A. 1989 Oct;86(20):8108-12 PMID: 2510160
  7. DNA related to the transforming gene(s) of avian sarcoma viruses is present in normal avian DNA.
    Nature. 1976 Mar 11;260(5547):170-3 PMID: 176594
  8. Proteins of synaptic vesicles involved in exocytosis and membrane recycling.
    Neuron. 1991 May;6(5):665-77 PMID: 1673848
  9. The neural network underlying locomotion in lamprey--synaptic and cellular mechanisms.
    Neuron. 1991 Jul;7(1):1-15 PMID: 1676892
  10. The src protein contains multiple domains for specific attachment to membranes.
    Mol Cell Biol. 1990 Mar;10(3):1000-9 PMID: 1689455
  11. Synaptophysin: a substrate for the protein tyrosine kinase pp60c-src in intact synaptic vesicles.
    Oncogene. 1990 Jul;5(7):1019-24 PMID: 1695724
  12. Intracellular localization of pp60c-src in human platelets.
    Oncogene. 1990 Jul;5(7):1033-6 PMID: 1695725
  13. Immunolocalization of the cellular src protein in interphase and mitotic NIH c-src overexpresser cells.
    J Cell Biol. 1990 Dec;111(6 Pt 2):3097-116 PMID: 1702788
  14. Specific proto-oncogenic tyrosine kinases of src family are enriched in cell-to-cell adherens junctions where the level of tyrosine phosphorylation is elevated.
    J Cell Biol. 1991 May;113(4):867-79 PMID: 1709169
  15. Identification of platelet membrane proteins that interact with amino-terminal peptides of pp60c-src.
    J Biol Chem. 1991 Oct 5;266(28):19040-6 PMID: 1717459
  16. A small GTP-binding protein dissociates from synaptic vesicles during exocytosis.
    Nature. 1991 Jan 3;349(6304):79-81 PMID: 1845915
  17. Redistribution of synaptophysin and synapsin I during alpha-latrotoxin-induced release of neurotransmitter at the neuromuscular junction.
    J Cell Biol. 1990 Feb;110(2):449-59 PMID: 1967610
  18. Targeted disruption of the c-src proto-oncogene leads to osteopetrosis in mice.
    Cell. 1991 Feb 22;64(4):693-702 PMID: 1997203
  19. Biogenesis of synaptic vesicle-like structures in a pheochromocytoma cell line PC-12.
    J Cell Biol. 1990 May;110(5):1693-703 PMID: 2110571
  20. Localization and subcellular distribution of smg p25A, a ras p21-like GTP-binding protein, in rat brain.
    J Biol Chem. 1990 Jul 15;265(20):11872-9 PMID: 2114404
  21. rab3 is a small GTP-binding protein exclusively localized to synaptic vesicles.
    Proc Natl Acad Sci U S A. 1990 Mar;87(5):1988-92 PMID: 2155429
  22. Identification of a 32K plasma membrane protein that binds to the myristylated amino-terminal sequence of p60v-src.
    Nature. 1990 Jul 5;346(6279):84-6 PMID: 2164157
  23. Neurones express high levels of a structurally modified, activated form of pp60c-src.
    Nature. 1985 Aug 8-14;316(6028):554-7 PMID: 2412121
  24. Synaptophysin is targeted to similar microvesicles in CHO and PC12 cells.
    EMBO J. 1989 Oct;8(10):2863-72 PMID: 2510994
  25. Specific and saturable binding of pp60v-src to plasma membranes: evidence for a myristyl-src receptor.
    Cell. 1989 Jul 28;58(2):281-6 PMID: 2546680
  26. Highly specific antibody to Rous sarcoma virus src gene product recognizes a novel population of pp60v-src and pp60c-src molecules.
    J Cell Biol. 1985 Feb;100(2):409-17 PMID: 2981886
  27. Mutation of NH2-terminal glycine of p60src prevents both myristoylation and morphological transformation.
    Proc Natl Acad Sci U S A. 1985 Jul;82(14):4625-8 PMID: 2991884
  28. The absence of myristic acid decreases membrane binding of p60src but does not affect tyrosine protein kinase activity.
    J Virol. 1986 May;58(2):468-74 PMID: 3009860
  29. Processing of p60v-src to its myristylated membrane-bound form.
    Mol Cell Biol. 1985 Oct;5(10):2781-8 PMID: 3016513
  30. Characterization of synapsin I binding to small synaptic vesicles.
    J Biol Chem. 1986 Jun 25;261(18):8383-90 PMID: 3087973
  31. Protein p38: an integral membrane protein specific for small vesicles of neurons and neuroendocrine cells.
    J Cell Biol. 1986 Dec;103(6 Pt 1):2511-27 PMID: 3097029
  32. Fractionation of synaptophysin-containing vesicles from rat brain and cultured PC12 pheochromocytoma cells.
    FEBS Lett. 1988 Nov 21;240(1-2):71-7 PMID: 3142795
  33. Identification, by a monoclonal antibody, of a 53-kD protein associated with a tubulo-vesicular compartment at the cis-side of the Golgi apparatus.
    J Cell Biol. 1988 Nov;107(5):1643-53 PMID: 3182932
  34. p60c-src activity detected in the chromaffin granule membrane.
    Biochem Biophys Res Commun. 1986 Jan 29;134(2):736-42 PMID: 3511908
  35. A short sequence in the p60src N terminus is required for p60src myristylation and membrane association and for cell transformation.
    Mol Cell Biol. 1984 Sep;4(9):1834-42 PMID: 6092942
  36. Neural tissues express high levels of the cellular src gene product pp60c-src.
    Mol Cell Biol. 1983 Jun;3(6):1157-62 PMID: 6192323
  37. The protein encoded by the transforming gene of avian sarcoma virus (pp60src) and a homologous protein in normal cells (pp60proto-src) are associated with the plasma membrane.
    Proc Natl Acad Sci U S A. 1980 Jul;77(7):3783-7 PMID: 6253989
  38. Phase separation of integral membrane proteins in Triton X-114 solution.
    J Biol Chem. 1981 Feb 25;256(4):1604-7 PMID: 6257680
  39. Changes in amino-terminal sequences of pp60src lead to decreased membrane association and decreased in vivo tumorigenicity.
    Cell. 1982 Apr;28(4):889-96 PMID: 6284379
  40. The cellular src gene product regulates junctional cell-to-cell communication.
    Science. 1988 Jan 22;239(4838):398-401 PMID: 2447651
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.117.5.1077
Published
1992-06-00
Pages
1077-84
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2289481
Subset
IM
Grants
NIDDK NIH HHS · DK33937 · United States
NINDS NIH HHS · NS09878 · United States
NINDS NIH HHS · NS15927 · United States
External Links
PubMed source DOI: 10.1083/jcb.117.5.1077 PMC Full Text View journal details
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