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PMID: 7689149 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Suppression of c-Src activity by C-terminal Src kinase involves the c-Src SH2 and SH3 domains: analysis with Saccharomyces cerevisiae.

Molecular and cellular biology ·Vol. 13 ·No. 9 ·1993-09-00 ·Pages 5290-300

Murphy SM, Bergman M, Morgan DO

Abstract

The kinase activity of c-Src is normally repressed in vertebrate cells by extensive phosphorylation of Y-527. C-terminal Src kinase (CSK) is a candidate for the enzyme that catalyzes this phosphorylation. We have used budding yeast to study the regulation of c-Src activity by CSK in intact cells. Expression of c-Src in Saccharomyces cerevisiae, which lacks endogenous c-Src and Y-527 kinases, induces a kinase-dependent growth inhibition. Coexpression of CSK in these cells results in phosphorylation of c-Src on Y-527 and suppression of the c-Src phenotype. CSK does not fully suppress the activity of c-Src mutants lacking portions of the SH2 or SH3 domains, even though these mutant proteins are phosphorylated on Y-527 by CSK both in vivo and in vitro. These results suggest that both the SH2 and SH3 domains of c-Src are required for the suppression of c-Src activity by Y-527 phosphorylation.

MeSH Terms
CSK Tyrosine-Protein Kinase Cell Division DNA Mutational Analysis Oncogene Protein pp60(v-src)/metabolism Phenotype Phosphoproteins/metabolism Phosphorylation Phosphotyrosine Protein-Tyrosine Kinases/metabolism Proto-Oncogene Proteins pp60(c-src)/metabolism Recombinant Fusion Proteins/metabolism Saccharomyces cerevisiae Structure-Activity Relationship Tyrosine/analogs & derivatives,metabolism src-Family Kinases
Chemicals
Phosphoproteins Recombinant Fusion Proteins Phosphotyrosine Tyrosine Protein-Tyrosine Kinases CSK Tyrosine-Protein Kinase Oncogene Protein pp60(v-src) Proto-Oncogene Proteins pp60(c-src) src-Family Kinases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Murphy S M
Department of Physiology, University of California, San Francisco 94143-0444.
Bergman M
Morgan D O
References (50)
50 references, click to expand
  1. Binding of transforming protein, P47gag-crk, to a broad range of phosphotyrosine-containing proteins.
    Science. 1990 Jun 22;248(4962):1537-9 PMID: 1694307
  2. Cloning of three human multifunctional de novo purine biosynthetic genes by functional complementation of yeast mutations.
    Proc Natl Acad Sci U S A. 1990 Apr;87(8):2916-20 PMID: 2183217
  3. 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
  4. Targeted disruption of the c-src proto-oncogene leads to osteopetrosis in mice.
    Cell. 1991 Feb 22;64(4):693-702 PMID: 1997203
  5. Getting started with yeast.
    Methods Enzymol. 1991;194:3-21 PMID: 2005794
  6. Cloning of a complementary DNA for a protein-tyrosine kinase that specifically phosphorylates a negative regulatory site of p60c-src.
    Nature. 1991 May 2;351(6321):69-72 PMID: 1709258
  7. cyl encodes a putative cytoplasmic tyrosine kinase lacking the conserved tyrosine autophosphorylation site (Y416src).
    Oncogene. 1991 Nov;6(11):2013-8 PMID: 1945408
  8. Phosphorylation of c-Src on tyrosine 527 by another protein tyrosine kinase.
    Science. 1991 Oct 25;254(5031):568-71 PMID: 1719633
  9. Design and use of peptide substrates for protein kinases.
    Methods Enzymol. 1991;200:121-34 PMID: 1835511
  10. Production of p60c-src by baculovirus expression and immunoaffinity purification.
    Methods Enzymol. 1991;200:645-60 PMID: 1720191
  11. 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
  12. CSK: a protein-tyrosine kinase involved in regulation of src family kinases.
    J Biol Chem. 1991 Dec 25;266(36):24249-52 PMID: 1722201
  13. Effects of SH2 and SH3 deletions on the functional activities of wild-type and transforming variants of c-Src.
    Mol Cell Biol. 1992 Apr;12(4):1835-45 PMID: 1549129
  14. 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
  15. Association of p60c-src with endosomal membranes in mammalian fibroblasts.
    J Cell Biol. 1992 Jul;118(2):321-33 PMID: 1378446
  16. The human p50csk tyrosine kinase phosphorylates p56lck at Tyr-505 and down regulates its catalytic activity.
    EMBO J. 1992 Aug;11(8):2919-24 PMID: 1639064
  17. SH3--an abundant protein domain in search of a function.
    FEBS Lett. 1992 Jul 27;307(1):55-61 PMID: 1639195
  18. Crystal structure of the phosphotyrosine recognition domain SH2 of v-src complexed with tyrosine-phosphorylated peptides.
    Nature. 1992 Aug 20;358(6388):646-53 PMID: 1379696
  19. Identification of a protein that binds to the SH3 region of Abl and is similar to Bcr and GAP-rho.
    Science. 1992 Aug 7;257(5071):803-6 PMID: 1379745
  20. SH2 and SH3 domains: from structure to function.
    Cell. 1992 Oct 30;71(3):359-62 PMID: 1423600
  21. Solution structure of the SH3 domain of Src and identification of its ligand-binding site.
    Science. 1992 Dec 4;258(5088):1665-8 PMID: 1280858
  22. Platelet activation leads to increased c-src kinase activity and association of c-src with an 85-kDa tyrosine phosphoprotein.
    Oncogene. 1992 Dec;7(12):2407-15 PMID: 1281303
  23. Phosphorylation of Src mutants at Tyr 527 in fibroblasts does not correlate with in vitro phosphorylation by CSK.
    Oncogene. 1993 Jan;8(1):117-24 PMID: 7678701
  24. Redistribution of activated pp60c-src to integrin-dependent cytoskeletal complexes in thrombin-stimulated platelets.
    Mol Cell Biol. 1993 Mar;13(3):1863-71 PMID: 7680100
  25. The cell cycle and c-Src.
    Curr Opin Genet Dev. 1993 Feb;3(1):26-34 PMID: 7680927
  26. Osteopetrosis in Src-deficient mice is due to an autonomous defect of osteoclasts.
    Proc Natl Acad Sci U S A. 1993 May 15;90(10):4485-9 PMID: 7685105
  27. Preparation of RNA and ribosomes from yeast.
    Methods Cell Biol. 1975;12:45-64 PMID: 1105073
  28. Corrections to the nucleotide sequence of the src gene of Rous sarcoma virus.
    Nature. 1983 Feb 24;301(5902):736-8 PMID: 6298633
  29. Isolation of monoclonal antibodies that recognize the transforming proteins of avian sarcoma viruses.
    J Virol. 1983 Nov;48(2):352-60 PMID: 6312092
  30. Monoclonal antibodies to Rous sarcoma virus pp60src react with enzymatically active cellular pp60src of avian and mammalian origin.
    J Virol. 1984 Aug;51(2):272-82 PMID: 6205164
  31. p60c-src activity detected in the chromaffin granule membrane.
    Biochem Biophys Res Commun. 1986 Jan 29;134(2):736-42 PMID: 3511908
  32. Tyr527 is phosphorylated in pp60c-src: implications for regulation.
    Science. 1986 Mar 21;231(4744):1431-4 PMID: 2420005
  33. Amino acid substitutions sufficient to convert the nontransforming p60c-src protein to a transforming protein.
    Mol Cell Biol. 1986 Dec;6(12):4155-60 PMID: 2432397
  34. Dephosphorylation or antibody binding to the carboxy terminus stimulates pp60c-src.
    Mol Cell Biol. 1986 Dec;6(12):4467-77 PMID: 2432403
  35. 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
  36. Tyrosine phosphorylation regulates the biochemical and biological properties of pp60c-src.
    Cell. 1987 Apr 10;49(1):75-82 PMID: 3103926
  37. Cell transformation by pp60c-src mutated in the carboxy-terminal regulatory domain.
    Cell. 1987 Apr 10;49(1):83-91 PMID: 3103927
  38. Characterization of avian and viral p60src proteins expressed in yeast.
    Proc Natl Acad Sci U S A. 1987 Jul;84(13):4455-9 PMID: 2440023
  39. Activation of the oncogenic potential of the avian cellular src protein by specific structural alteration of the carboxy terminus.
    EMBO J. 1987 Aug;6(8):2359-64 PMID: 2822389
  40. Avian pp60c-src is more active when expressed in yeast than in vertebrate fibroblasts.
    Oncogene Res. 1987 Sep-Oct;1(4):297-310 PMID: 2453017
  41. Investigation of factors that influence phosphorylation of pp60c-src on tyrosine 527.
    Mol Cell Biol. 1988 Jun;8(6):2465-71 PMID: 2457151
  42. Regulation by the autophosphorylation site in overexpressed pp60c-src.
    Mol Cell Biol. 1988 Oct;8(10):4541-6 PMID: 2460746
  43. 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
  44. Platelet-derived growth factor induces multisite phosphorylation of pp60c-src and increases its protein-tyrosine kinase activity.
    Mol Cell Biol. 1988 Aug;8(8):3345-56 PMID: 2463476
  45. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
    Genetics. 1989 May;122(1):19-27 PMID: 2659436
  46. 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
  47. A protein tyrosine kinase involved in regulation of pp60c-src function.
    J Biol Chem. 1989 Dec 15;264(35):20886-93 PMID: 2480346
  48. The src protein contains multiple domains for specific attachment to membranes.
    Mol Cell Biol. 1990 Mar;10(3):1000-9 PMID: 1689455
  49. 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
  50. Mutations in src homology regions 2 and 3 of activated chicken c-src that result in preferential transformation of mouse or chicken cells.
    Proc Natl Acad Sci U S A. 1990 Nov;87(21):8592-6 PMID: 1700434
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.13.9.5290-5300.1993
Published
1993-09-00
Pages
5290-300
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC360223
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.13.9.5290-5300.1993 PMC Full Text View journal details
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