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PMID: 7525268 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Association of the amino-terminal half of c-Src with focal adhesions alters their properties and is regulated by phosphorylation of tyrosine 527.

The EMBO journal ·Vol. 13 ·No. 20 ·1994-10-17 ·Pages 4745-56

Kaplan KB, Bibbins KB, Swedlow JR, Arnaud M, Morgan DO, Varmus HE

Abstract

We have characterized the mechanism by which the subcellular distribution of c-Src is controlled by the phosphorylation of tyrosine 527. Mutation of this tyrosine dramatically redistributes c-Src from endosomal membranes to focal adhesions. Redistribution to focal adhesions occurs independently of kinase activity and cellular transformation. In cells lacking the regulatory kinase (CSK) that phosphorylates tyrosine 527, c-Src is also found predominantly in focal adhesions, confirming that phosphorylation of tyrosine 527 affects the location of c-Src inside the cell. The first 251 amino acids of c-Src are sufficient to allow association with focal adhesions, indicating that at least one signal for positioning c-Src in focal adhesions resides in the amino-terminal half. Point mutations and deletions in the first 251 amino acids of c-Src reveal that association with focal adhesions requires the myristylation site needed for membrane attachment, as well as the SH3 domain. Expression of the amino-terminal region alters both the structural and biochemical properties of focal adhesions. Focal adhesions containing this non-catalytic portion of c-Src are larger and exhibit increased levels of phosphotyrosine staining. Our results suggest that c-Src may regulate focal adhesions and cellular adhesion by a kinase-independent mechanism.

MeSH Terms
Amino Acid Sequence Animals Biological Transport Cell Adhesion Molecules/chemistry,metabolism Cell Line Mice Molecular Sequence Data Phosphorylation Protein Binding Protein Sorting Signals/physiology Proto-Oncogene Proteins pp60(c-src)/chemistry,metabolism Structure-Activity Relationship Subcellular Fractions/metabolism Tyrosine/metabolism
Chemicals
Cell Adhesion Molecules Protein Sorting Signals Tyrosine Proto-Oncogene Proteins pp60(c-src)
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Kaplan K B
Department of Microbiology and Immunology, University of California, San Francisco 94143.
Bibbins K B
Swedlow J R
Arnaud M
Morgan D O
Varmus H E
References (48)
48 references, click to expand
  1. SH2 and SH3 domains: from structure to function.
    Cell. 1992 Oct 30;71(3):359-62 PMID: 1423600
  2. Localization of the viral and cellular Src kinases to perinuclear vesicles in fibroblasts.
    Cell Growth Differ. 1992 Sep;3(9):567-76 PMID: 1384654
  3. Regulation of c-Src tyrosine kinase activity by the Src SH2 domain.
    Oncogene. 1993 May;8(5):1119-26 PMID: 7683128
  4. Disruption of the csk gene, encoding a negative regulator of Src family tyrosine kinases, leads to neural tube defects and embryonic lethality in mice.
    Cell. 1993 Jun 18;73(6):1117-24 PMID: 7685657
  5. Detection of Src homology 3-binding proteins, including paxillin, in normal and v-Src-transformed Balb/c 3T3 cells.
    J Biol Chem. 1993 Jul 15;268(20):14956-63 PMID: 8325872
  6. Csk inhibition of c-Src activity requires both the SH2 and SH3 domains of Src.
    EMBO J. 1993 Jul;12(7):2625-34 PMID: 7687537
  7. The v-Src SH3 domain binds phosphatidylinositol 3'-kinase.
    Mol Cell Biol. 1993 Sep;13(9):5225-32 PMID: 7689147
  8. Association of the src gene product of Rous sarcoma virus with cytoskeletal structures of chicken embryo fibroblasts.
    Proc Natl Acad Sci U S A. 1980 Jun;77(6):3484-8 PMID: 6251461
  9. Adhesion plaques of Rous sarcoma virus-transformed cells contain the src gene product.
    Proc Natl Acad Sci U S A. 1980 Jun;77(6):3514-8 PMID: 6251464
  10. Altered distributions of the cytoskeletal proteins vinculin and alpha-actinin in cultured fibroblasts transformed by Rous sarcoma virus.
    Proc Natl Acad Sci U S A. 1980 Nov;77(11):6687-91 PMID: 6256755
  11. Immunofluorescent localization of the transforming protein of Rous sarcoma virus with antibodies against a synthetic src peptide.
    Proc Natl Acad Sci U S A. 1982 Sep;79(17):5322-6 PMID: 6291037
  12. Neural tissues express high levels of the cellular src gene product pp60c-src.
    Mol Cell Biol. 1983 Jun;3(6):1157-62 PMID: 6192323
  13. Myristic acid, a rare fatty acid, is the lipid attached to the transforming protein of Rous sarcoma virus and its cellular homolog.
    J Virol. 1985 Jan;53(1):7-12 PMID: 2981363
  14. 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
  15. Activation of the pp60c-src kinase by middle T antigen binding or by dephosphorylation.
    EMBO J. 1985 Jun;4(6):1471-7 PMID: 2411538
  16. The product of the protooncogene c-src is modified during the cellular response to platelet-derived growth factor.
    Proc Natl Acad Sci U S A. 1985 Dec;82(23):7845-9 PMID: 2415973
  17. Blood platelets express high levels of the pp60c-src-specific tyrosine kinase activity.
    Proc Natl Acad Sci U S A. 1986 Feb;83(4):852-6 PMID: 2419901
  18. Activation and suppression of pp60c-src transforming ability by mutation of its primary sites of tyrosine phosphorylation.
    Cell. 1987 Apr 10;49(1):65-73 PMID: 3103925
  19. Tyrosine phosphorylation regulates the biochemical and biological properties of pp60c-src.
    Cell. 1987 Apr 10;49(1):75-82 PMID: 3103926
  20. Association of p60src with Triton X-100-resistant cellular structure correlates with morphological transformation.
    Proc Natl Acad Sci U S A. 1987 Apr;84(8):2312-6 PMID: 3031677
  21. Altered phosphorylation and activation of pp60c-src during fibroblast mitosis.
    Cell. 1988 Mar 25;52(6):801-10 PMID: 2450676
  22. Regulation by the autophosphorylation site in overexpressed pp60c-src.
    Mol Cell Biol. 1988 Oct;8(10):4541-6 PMID: 2460746
  23. 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
  24. Fluorescence microscopy in three dimensions.
    Methods Cell Biol. 1989;30:353-77 PMID: 2494418
  25. Mitosis-specific phosphorylation of p60c-src by p34cdc2-associated protein kinase.
    Cell. 1989 Jun 2;57(5):775-86 PMID: 2470513
  26. Structural differences between repressed and derepressed forms of p60c-src.
    Mol Cell Biol. 1989 Jun;9(6):2648-56 PMID: 2474758
  27. The src protein contains multiple domains for specific attachment to membranes.
    Mol Cell Biol. 1990 Mar;10(3):1000-9 PMID: 1689455
  28. Site-directed mutagenesis of the SH2- and SH3-coding domains of c-src produces varied phenotypes, including oncogenic activation of p60c-src.
    Mol Cell Biol. 1990 Apr;10(4):1307-18 PMID: 2108315
  29. Association between the PDGF receptor and members of the src family of tyrosine kinases.
    Cell. 1990 Aug 10;62(3):481-92 PMID: 1696179
  30. Paxillin: a new vinculin-binding protein present in focal adhesions.
    J Cell Biol. 1990 Sep;111(3):1059-68 PMID: 2118142
  31. 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
  32. Suppression of c-Src activity by C-terminal Src kinase involves the c-Src SH2 and SH3 domains: analysis with Saccharomyces cerevisiae.
    Mol Cell Biol. 1993 Sep;13(9):5290-300 PMID: 7689149
  33. The GTPase dynamin binds to and is activated by a subset of SH3 domains.
    Cell. 1993 Oct 8;75(1):25-36 PMID: 8402898
  34. Binding of the Src SH2 domain to phosphopeptides is determined by residues in both the SH2 domain and the phosphopeptides.
    Mol Cell Biol. 1993 Dec;13(12):7278-87 PMID: 7504171
  35. Caveolae: where incoming and outgoing messengers meet.
    Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):10909-13 PMID: 8248193
  36. Stable association of pp60src and pp59fyn with the focal adhesion-associated protein tyrosine kinase, pp125FAK.
    Mol Cell Biol. 1994 Jan;14(1):147-55 PMID: 7505391
  37. The noncatalytic src homology region 2 segment of abl tyrosine kinase binds to tyrosine-phosphorylated cellular proteins with high affinity.
    Proc Natl Acad Sci U S A. 1991 Jan 15;88(2):627-31 PMID: 1703304
  38. Deletions in the SH2 domain of p60v-src prevent association with the detergent-insoluble cellular matrix.
    Mol Cell Biol. 1991 Mar;11(3):1207-13 PMID: 1705002
  39. Targeted disruption of the c-src proto-oncogene leads to osteopetrosis in mice.
    Cell. 1991 Feb 22;64(4):693-702 PMID: 1997203
  40. 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
  41. Dominant positive and negative selection using a hygromycin phosphotransferase-thymidine kinase fusion gene.
    Mol Cell Biol. 1991 Jun;11(6):3374-8 PMID: 1645450
  42. Three-dimensional multiple-wavelength fluorescence microscopy for the structural analysis of biological phenomena.
    Semin Cell Biol. 1991 Jun;2(3):153-65 PMID: 1720334
  43. Selective binding of activated pp60c-src by an immobilized synthetic phosphopeptide modeled on the carboxyl terminus of pp60c-src.
    Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10696-700 PMID: 1720546
  44. Point mutations in the abl SH2 domain coordinately impair phosphotyrosine binding in vitro and transforming activity in vivo.
    Mol Cell Biol. 1992 Feb;12(2):609-18 PMID: 1370711
  45. Specific association of the proto-oncogene product pp60c-src with an intracellular organelle, the PC12 synaptic vesicle.
    J Cell Biol. 1992 Jun;117(5):1077-84 PMID: 1374414
  46. Association of p60c-src with endosomal membranes in mammalian fibroblasts.
    J Cell Biol. 1992 Jul;118(2):321-33 PMID: 1378446
  47. Requirement of pp60c-src expression for osteoclasts to form ruffled borders and resorb bone in mice.
    J Clin Invest. 1992 Oct;90(4):1622-7 PMID: 1383278
  48. Identification of a ten-amino acid proline-rich SH3 binding site.
    Science. 1993 Feb 19;259(5098):1157-61 PMID: 8438166
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1994-10-17
Pages
4745-56
Language
English
Region
England
NLM ID
8208664
PMCID
PMC395413
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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