Home LiteratureArticle Details
PMID: 8227154 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Identification of sequences required for the efficient localization of the focal adhesion kinase, pp125FAK, to cellular focal adhesions.

The Journal of cell biology ·Vol. 123 ·No. 4 ·1993-11-00 ·Pages 993-1005

Hildebrand JD, Schaller MD, Parsons JT

Abstract

The integrin family of heterodimeric cell surface receptors play critical roles in multiple biological processes by mediating cellular adhesion to the extracellular matrix (ECM). Adhesion triggers intracellular signaling cascades, including tyrosine phosphorylation and elevation of [Ca2+]i. The Focal Adhesion Kinase (FAK or pp125FAK), a protein tyrosine kinase that colocalizes with integrins in cellular focal adhesions, is a prime candidate for a mediator of integrin signaling events. Here we report an analysis of the domain structure of FAK in which we have identified a contiguous stretch of 159 amino acids within the COOH terminus essential for correct subcellular localization. When placed in the context of an unrelated cytosolic protein, this Focal Adhesion Targeting (FAT) sequence functions to efficiently mediate the focal adhesion localization of this fusion protein. Furthermore, this analysis suggests that pp125FAK cannot be activated oncogenically by mutation. This result could be explained if pp125FK either exhibits a narrow substrate specificity or is diametrically opposed by cellular phosphatases or other cellular processes.

MeSH Terms
Amino Acid Sequence Animals Cell Adhesion Cell Adhesion Molecules/metabolism Cell Line Cells, Cultured Chick Embryo Extracellular Matrix/metabolism Fluorescent Antibody Technique Focal Adhesion Kinase 1 Focal Adhesion Protein-Tyrosine Kinases Molecular Sequence Data Peptide Fragments/metabolism Phosphorylation Protein-Tyrosine Kinases/metabolism Receptor, Insulin/metabolism Signal Transduction Transfection
Chemicals
Cell Adhesion Molecules Peptide Fragments Protein-Tyrosine Kinases Receptor, Insulin Focal Adhesion Kinase 1 Focal Adhesion Protein-Tyrosine Kinases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Hildebrand J D
Department of Microbiology, University of Virginia, Charlottesville 22908.
Schaller M D
Parsons J T
References (60)
60 references, click to expand
  1. Functional studies of the domains of talin.
    J Cell Biol. 1990 May;110(5):1635-44 PMID: 2110569
  2. Two novel protein-tyrosine kinases, each with a second phosphotransferase-related catalytic domain, define a new class of protein kinase.
    Mol Cell Biol. 1991 Apr;11(4):2057-65 PMID: 1848670
  3. Focal adhesion kinase: an integrin-linked protein tyrosine kinase.
    Trends Cell Biol. 1993 Aug;3(8):258-62 PMID: 14731743
  4. Molecular cloning of a porcine gene syk that encodes a 72-kDa protein-tyrosine kinase showing high susceptibility to proteolysis.
    J Biol Chem. 1991 Aug 25;266(24):15790-6 PMID: 1874735
  5. Autonomous expression of a noncatalytic domain of the focal adhesion-associated protein tyrosine kinase pp125FAK.
    Mol Cell Biol. 1993 Feb;13(2):785-91 PMID: 8423801
  6. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  7. Role of platelet membrane glycoprotein IIb-IIIa in agonist-induced tyrosine phosphorylation of platelet proteins.
    J Cell Biol. 1990 Dec;111(6 Pt 2):3117-27 PMID: 1702789
  8. Monoclonal antibodies specific for the carboxy terminus of simian virus 40 large T antigen.
    J Virol. 1984 Nov;52(2):483-91 PMID: 6208378
  9. DNA sequence, structure, and tyrosine kinase activity of the Drosophila melanogaster Abelson proto-oncogene homolog.
    Mol Cell Biol. 1988 Feb;8(2):843-53 PMID: 2832740
  10. Transmembrane molecular assemblies in cell-extracellular matrix interactions.
    Curr Opin Cell Biol. 1991 Oct;3(5):849-53 PMID: 1931085
  11. Signal transduction through the fibronectin receptor induces collagenase and stromelysin gene expression.
    J Cell Biol. 1989 Aug;109(2):877-89 PMID: 2547805
  12. Tyrosine phosphorylation of paxillin and pp125FAK accompanies cell adhesion to extracellular matrix: a role in cytoskeletal assembly.
    J Cell Biol. 1992 Nov;119(4):893-903 PMID: 1385444
  13. Cell adhesion or integrin clustering increases phosphorylation of a focal adhesion-associated tyrosine kinase.
    J Biol Chem. 1992 Nov 25;267(33):23439-42 PMID: 1429685
  14. 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
  15. Signal transduction by receptors with tyrosine kinase activity.
    Cell. 1990 Apr 20;61(2):203-12 PMID: 2158859
  16. tyk2, prototype of a novel class of non-receptor tyrosine kinase genes.
    Oncogene. 1990 Sep;5(9):1329-36 PMID: 2216457
  17. Interaction of plasma membrane fibronectin receptor with talin--a transmembrane linkage.
    Nature. 1986 Apr 10-16;320(6062):531-3 PMID: 2938015
  18. Oncogenes and signal transduction.
    Cell. 1991 Jan 25;64(2):281-302 PMID: 1846320
  19. pp125FAK a structurally distinctive protein-tyrosine kinase associated with focal adhesions.
    Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):5192-6 PMID: 1594631
  20. Integrins isolated from Rous sarcoma virus-transformed chicken embryo fibroblasts.
    Oncogene. 1989 Mar;4(3):325-33 PMID: 2468126
  21. Regulation of focal adhesion-associated protein tyrosine kinase by both cellular adhesion and oncogenic transformation.
    Nature. 1992 Aug 20;358(6388):690-2 PMID: 1379699
  22. Spreading of human endothelial cells on fibronectin or vitronectin triggers elevation of intracellular free calcium.
    J Cell Biol. 1993 Feb;120(4):1003-10 PMID: 7679387
  23. Presence of an SH2 domain in the actin-binding protein tensin.
    Science. 1991 May 3;252(5006):712-5 PMID: 1708917
  24. Zyxin and cCRP: two interactive LIM domain proteins associated with the cytoskeleton.
    J Cell Biol. 1992 Dec;119(6):1573-87 PMID: 1469049
  25. Tyrosine phosphorylation of the focal adhesion kinase pp125FAK during development: relation to paxillin.
    J Cell Sci. 1993 Jul;105 ( Pt 3):637-45 PMID: 8408291
  26. A protein tyrosine kinase in the interferon alpha/beta signaling pathway.
    Cell. 1992 Jul 24;70(2):313-22 PMID: 1386289
  27. Multiple elevations of cytosolic-free Ca2+ in human neutrophils: initiation by adherence receptors of the integrin family.
    J Cell Biol. 1991 Mar;112(6):1249-57 PMID: 1900302
  28. Neonatal lethality and lymphopenia in mice with a homozygous disruption of the c-abl proto-oncogene.
    Cell. 1991 Jun 28;65(7):1153-63 PMID: 2065352
  29. Intracellular targeting of pp60src expression: localization of v-src to adhesion plaques is sufficient to transform chicken embryo fibroblasts.
    Oncogene. 1992 Dec;7(12):2417-28 PMID: 1334249
  30. Signal transduction by integrins: increased protein tyrosine phosphorylation caused by clustering of beta 1 integrins.
    Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8392-6 PMID: 1717976
  31. The when and how of Src regulation.
    Cell. 1993 Jun 18;73(6):1051-4 PMID: 7685656
  32. B lymphocyte activation is accompanied by phosphorylation of a 72-kDa protein-tyrosine kinase.
    J Biol Chem. 1991 Aug 15;266(23):14846-9 PMID: 1869521
  33. Myristylation is required for Tyr-527 dephosphorylation and activation of pp60c-src in mitosis.
    Mol Cell Biol. 1993 Mar;13(3):1464-70 PMID: 7680096
  34. The protein kinase family: conserved features and deduced phylogeny of the catalytic domains.
    Science. 1988 Jul 1;241(4861):42-52 PMID: 3291115
  35. Transformation-specific tyrosine phosphorylation of a novel cellular protein in chicken cells expressing oncogenic variants of the avian cellular src gene.
    Mol Cell Biol. 1989 Feb;9(2):629-38 PMID: 2469003
  36. Genetics of src: structure and functional organization of a protein tyrosine kinase.
    Curr Top Microbiol Immunol. 1989;147:79-127 PMID: 2482802
  37. Focal adhesion protein-tyrosine kinase phosphorylated in response to cell attachment to fibronectin.
    Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8487-91 PMID: 1528852
  38. Monoclonal antibodies to individual tyrosine-phosphorylated protein substrates of oncogene-encoded tyrosine kinases.
    Proc Natl Acad Sci U S A. 1990 May;87(9):3328-32 PMID: 2110361
  39. T cell antigen receptor signal transduction: a tale of tails and cytoplasmic protein-tyrosine kinases.
    Cell. 1993 Apr 23;73(2):209-12 PMID: 8477442
  40. Disruption of the actin cytoskeleton after microinjection of proteolytic fragments of alpha-actinin.
    J Cell Biol. 1991 Aug;114(3):481-91 PMID: 1907287
  41. Fibronectin/integrin interaction induces tyrosine phosphorylation of a 120-kDa protein.
    Cell Regul. 1991 Nov;2(11):951-64 PMID: 1725602
  42. Identification of a talin binding site in the cytoskeletal protein vinculin.
    J Cell Biol. 1989 Dec;109(6 Pt 1):2917-27 PMID: 2512301
  43. Focal adhesions: transmembrane junctions between the extracellular matrix and the cytoskeleton.
    Annu Rev Cell Biol. 1988;4:487-525 PMID: 3058164
  44. Crystal structure of the catalytic subunit of cyclic adenosine monophosphate-dependent protein kinase.
    Science. 1991 Jul 26;253(5018):407-14 PMID: 1862342
  45. Integrins and other cell adhesion molecules.
    FASEB J. 1990 Aug;4(11):2868-80 PMID: 2199285
  46. Tumor necrosis factor-induced degranulation in adherent human neutrophils is dependent on CD11b/CD18-integrin-triggered oscillations of cytosolic free Ca2+.
    Proc Natl Acad Sci U S A. 1990 Dec;87(23):9472-6 PMID: 1979172
  47. Signal transduction by the platelet integrin alpha IIb beta 3: induction of calcium oscillations required for protein-tyrosine phosphorylation and ligand-induced spreading of stably transfected cells.
    Mol Biol Cell. 1992 Sep;3(9):989-98 PMID: 1421580
  48. High-expression vectors with multiple cloning sites for construction of trpE fusion genes: pATH vectors.
    Methods Enzymol. 1991;194:477-90 PMID: 2005804
  49. Expression of an N-terminally truncated form of human focal adhesion kinase in brain.
    Biochem Biophys Res Commun. 1993 Jan 15;190(1):140-7 PMID: 8422239
  50. SH2 and SH3 domains: from structure to function.
    Cell. 1992 Oct 30;71(3):359-62 PMID: 1423600
  51. Identification and characterization of a novel cytoskeleton-associated pp60src substrate.
    Mol Cell Biol. 1991 Oct;11(10):5113-24 PMID: 1922035
  52. Integrins: versatility, modulation, and signaling in cell adhesion.
    Cell. 1992 Apr 3;69(1):11-25 PMID: 1555235
  53. An interaction between alpha-actinin and the beta 1 integrin subunit in vitro.
    J Cell Biol. 1990 Aug;111(2):721-9 PMID: 2116421
  54. Mice homozygous for the ablm1 mutation show poor viability and depletion of selected B and T cell populations.
    Cell. 1991 Jun 28;65(7):1165-75 PMID: 2065353
  55. Immunoaffinity purification of tyrosine-phosphorylated cellular proteins.
    J Immunol Methods. 1989 Jun 2;120(1):115-24 PMID: 2471744
  56. An actin-binding function contributes to transformation by the Bcr-Abl oncoprotein of Philadelphia chromosome-positive human leukemias.
    EMBO J. 1993 Apr;12(4):1533-46 PMID: 8467803
  57. Signal transduction from the extracellular matrix.
    J Cell Biol. 1993 Feb;120(3):577-85 PMID: 8381117
  58. Amino acid alterations within a highly conserved region of the Rous sarcoma virus src gene product pp60src inactivate tyrosine protein kinase activity.
    Mol Cell Biol. 1984 May;4(5):862-6 PMID: 6328273
  59. Adaptor plasmids simplify the insertion of foreign DNA into helper-independent retroviral vectors.
    J Virol. 1987 Oct;61(10):3004-12 PMID: 3041020
  60. Close similarity of epidermal growth factor receptor and v-erb-B oncogene protein sequences.
    Nature. 1984 Feb 9-15;307(5951):521-7 PMID: 6320011
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.123.4.993
Published
1993-11-00
Pages
993-1005
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2200138
Subset
IM
Grants
NCI NIH HHS · CA 29243 · United States
NCI NIH HHS · CA 40042 · United States
External Links
PubMed source DOI: 10.1083/jcb.123.4.993 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2