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J Cell Biol. 1993 May 2; 121(4): 909–921.
PMCID: PMC2119802

Expression of chicken vinculin complements the adhesion-defective phenotype of a mutant mouse F9 embryonal carcinoma cell


A mutant cell line, derived from the mouse embryonal carcinoma cell line F9, is defective in cell-cell adhesion (compaction) and in cell- substrate adhesion. We have previously shown that neither uvomorulin (E- cadherin) nor integrins are responsible for the mutant phenotype (Calogero, A., M. Samuels, T. Darland, S. A. Edwards, R. Kemler, and E. D. Adamson. 1991. Dev. Biol. 146:499-508). Several cytoskeleton proteins were assayed and only vinculin was found to be absent in mutant (5.51) cells. A chicken vinculin expression vector was transfected into the 5.51 cells together with a neomycin-resistance vector. Clones that were adherent to the substrate were selected in medium containing G418. Two clones, 5.51Vin3 and Vin4, were analyzed by Nomarski differential interference contrast and laser confocal microscopy as well as by biochemical and molecular biological techniques. Both clones adhered well to substrates and both exhibited F- actin stress fibers with vinculin localized at stress fiber tips in focal contacts. This was in marked contrast to 5.51 parental cells, which had no stress fibers and no vinculin. The mutant and complemented F9 cell lines will be useful models for examining the complex interactions between cytoskeletal and cell adhesion proteins.

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Selected References

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  • Adamson ED, Baribault H, Kemler R. Altered uvomorulin expression in a noncompacting mutant cell line of F9 embryonal carcinoma cells. Dev Biol. 1990 Apr;138(2):338–347. [PubMed]
  • Barstead RJ, Waterston RH. The basal component of the nematode dense-body is vinculin. J Biol Chem. 1989 Jun 15;264(17):10177–10185. [PubMed]
  • Bellas RE, Bendori R, Farmer SR. Epidermal growth factor activation of vinculin and beta 1-integrin gene transcription in quiescent Swiss 3T3 cells. Regulation through a protein kinase C-independent pathway. J Biol Chem. 1991 Jun 25;266(18):12008–12014. [PubMed]
  • Ben-Ze'ev A, Farmer SR, Penman S. Mechanisms of regulating tubulin synthesis in cultured mammalian cells. Cell. 1979 Jun;17(2):319–325. [PubMed]
  • Ben-Ze'ev A, Reiss R, Bendori R, Gorodecki B. Transient induction of vinculin gene expression in 3T3 fibroblasts stimulated by serum-growth factors. Cell Regul. 1990 Aug;1(9):621–636. [PMC free article] [PubMed]
  • Bendori R, Salomon D, Geiger B. Identification of two distinct functional domains on vinculin involved in its association with focal contacts. J Cell Biol. 1989 Jun;108(6):2383–2393. [PMC free article] [PubMed]
  • Black JD, Koury ST, Bankert RB, Repasky EA. Heterogeneity in lymphocyte spectrin distribution: ultrastructural identification of a new spectrin-rich cytoplasmic structure. J Cell Biol. 1988 Jan;106(1):97–109. [PMC free article] [PubMed]
  • Burridge K, Fath K, Kelly T, Nuckolls G, Turner C. Focal adhesions: transmembrane junctions between the extracellular matrix and the cytoskeleton. Annu Rev Cell Biol. 1988;4:487–525. [PubMed]
  • Burridge K, Feramisco JR. Microinjection and localization of a 130K protein in living fibroblasts: a relationship to actin and fibronectin. Cell. 1980 Mar;19(3):587–595. [PubMed]
  • Calogero A, Samuels M, Darland T, Edwards SA, Kemler R, Adamson ED. Overexpression of uvomorulin in a compaction-negative F9 mutant cell line. Dev Biol. 1991 Aug;146(2):499–508. [PubMed]
  • Chen WT, Singer SJ. Immunoelectron microscopic studies of the sites of cell-substratum and cell-cell contacts in cultured fibroblasts. J Cell Biol. 1982 Oct;95(1):205–222. [PMC free article] [PubMed]
  • Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. [PubMed]
  • Cleveland DW, Lopata MA, Sherline P, Kirschner MW. Unpolymerized tubulin modulates the level of tubulin mRNAs. Cell. 1981 Aug;25(2):537–546. [PubMed]
  • Cowin P, Kapprell HP, Franke WW, Tamkun J, Hynes RO. Plakoglobin: a protein common to different kinds of intercellular adhering junctions. Cell. 1986 Sep 26;46(7):1063–1073. [PubMed]
  • Critchley DR, Gilmore A, Hemmings L, Jackson P, McGregor A, Ohanian V, Patel B, Waites G, Wood C. Cytoskeletal proteins in adherens-type cell-matrix junctions. Biochem Soc Trans. 1991 Nov;19(4):1028–1033. [PubMed]
  • Darland T, Samuels M, Edwards SA, Sukhatme VP, Adamson ED. Regulation of Egr-1 (Zfp-6) and c-fos expression in differentiating embryonal carcinoma cells. Oncogene. 1991 Aug;6(8):1367–1376. [PubMed]
  • Dean DC, McQuillan JJ, Weintraub S. Serum stimulation of fibronectin gene expression appears to result from rapid serum-induced binding of nuclear proteins to a cAMP response element. J Biol Chem. 1990 Feb 25;265(6):3522–3527. [PubMed]
  • DePasquale JA, Izzard CS. Evidence for an actin-containing cytoplasmic precursor of the focal contact and the timing of incorporation of vinculin at the focal contact. J Cell Biol. 1987 Dec;105(6 Pt 1):2803–2809. [PMC free article] [PubMed]
  • DePasquale JA, Izzard CS. Accumulation of talin in nodes at the edge of the lamellipodium and separate incorporation into adhesion plaques at focal contacts in fibroblasts. J Cell Biol. 1991 Jun;113(6):1351–1359. [PMC free article] [PubMed]
  • Ezzell RM, Kenney DM, Egan S, Stossel TP, Hartwig JH. Localization of the domain of actin-binding protein that binds to membrane glycoprotein Ib and actin in human platelets. J Biol Chem. 1988 Sep 15;263(26):13303–13309. [PubMed]
  • Feinberg AP, Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. [PubMed]
  • Rodríguez Fernández JL, Geiger B, Salomon D, Ben-Ze'ev A. Overexpression of vinculin suppresses cell motility in BALB/c 3T3 cells. Cell Motil Cytoskeleton. 1992;22(2):127–134. [PubMed]
  • Rodríguez Fernández JL, Geiger B, Salomon D, Sabanay I, Zöller M, Ben-Ze'ev A. Suppression of tumorigenicity in transformed cells after transfection with vinculin cDNA. J Cell Biol. 1992 Oct;119(2):427–438. [PMC free article] [PubMed]
  • Geiger B. A 130K protein from chicken gizzard: its localization at the termini of microfilament bundles in cultured chicken cells. Cell. 1979 Sep;18(1):193–205. [PubMed]
  • Geiger B, Ginsberg D. The cytoplasmic domain of adherens-type junctions. Cell Motil Cytoskeleton. 1991;20(1):1–6. [PubMed]
  • Geiger B, Dutton AH, Tokuyasu KT, Singer SJ. Immunoelectron microscope studies of membrane-microfilament interactions: distributions of alpha-actinin, tropomyosin, and vinculin in intestinal epithelial brush border and chicken gizzard smooth muscle cells. J Cell Biol. 1981 Dec;91(3 Pt 1):614–628. [PMC free article] [PubMed]
  • Geiger B, Tokuyasu KT, Dutton AH, Singer SJ. Vinculin, an intracellular protein localized at specialized sites where microfilament bundles terminate at cell membranes. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4127–4131. [PubMed]
  • Graham FL, Abrahams PJ, Mulder C, Heijneker HL, Warnaar SO, De Vries FA, Fiers W, Van Der Eb AJ. Studies on in vitro transformation by DNA and DNA fragments of human adenoviruses and simian virus 40. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):637–650. [PubMed]
  • Grover A, Oshima RG, Adamson ED. Epithelial layer formation in differentiating aggregates of F9 embryonal carcinoma cells. J Cell Biol. 1983 Jun;96(6):1690–1696. [PMC free article] [PubMed]
  • Grover A, Rosentraus MJ, Sterman B, Snook ME, Adamson ED. An adhesion-defective variant of F9 embryonal carcinoma cells fails to differentiate into visceral endoderm. Dev Biol. 1987 Mar;120(1):1–11. [PubMed]
  • Hayashi Y, Haimovich B, Reszka A, Boettiger D, Horwitz A. Expression and function of chicken integrin beta 1 subunit and its cytoplasmic domain mutants in mouse NIH 3T3 cells. J Cell Biol. 1990 Jan;110(1):175–184. [PMC free article] [PubMed]
  • Herrenknecht K, Ozawa M, Eckerskorn C, Lottspeich F, Lenter M, Kemler R. The uvomorulin-anchorage protein alpha catenin is a vinculin homologue. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):9156–9160. [PubMed]
  • Horvath AR, Asijee GM, Muszbek L. Cytoskeletal assembly and vinculin-cytoskeleton interaction in different phases of the activation of bovine platelets. Cell Motil Cytoskeleton. 1992;21(2):123–131. [PubMed]
  • Hynes RO. Integrins: versatility, modulation, and signaling in cell adhesion. Cell. 1992 Apr 3;69(1):11–25. [PubMed]
  • Ingle CA, Drinkwater NR. Mutational specificities of 1'-acetoxysafrole, N-benzoyloxy-N-methyl-4-aminoazobenzene, and ethyl methanesulfonate in human cells. Mutat Res. 1989 Mar-May;220(2-3):133–142. [PubMed]
  • Jones P, Jackson P, Price GJ, Patel B, Ohanion V, Lear AL, Critchley DR. Identification of a talin binding site in the cytoskeletal protein vinculin. J Cell Biol. 1989 Dec;109(6 Pt 1):2917–2927. [PMC free article] [PubMed]
  • Kemler R, Ozawa M. Uvomorulin-catenin complex: cytoplasmic anchorage of a Ca2+-dependent cell adhesion molecule. Bioessays. 1989 Oct;11(4):88–91. [PubMed]
  • Koteliansky VE, Ogryzko EP, Zhidkova NI, Weller PA, Critchley DR, Vancompernolle K, Vandekerckhove J, Strasser P, Way M, Gimona M, et al. An additional exon in the human vinculin gene specifically encodes meta-vinculin-specific difference peptide. Cross-species comparison reveals variable and conserved motifs in the meta-vinculin insert. Eur J Biochem. 1992 Mar 1;204(2):767–772. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Lehtonen E, Lehto VP, Badley RA, Virtanen I. Formation of vinculin plaques precedes other cytoskeletal changes during retinoic acid-induced teratocarcinoma cell differentiation. Exp Cell Res. 1983 Mar;144(1):191–197. [PubMed]
  • Marcantonio EE, Guan JL, Trevithick JE, Hynes RO. Mapping of the functional determinants of the integrin beta 1 cytoplasmic domain by site-directed mutagenesis. Cell Regul. 1990 Jul;1(8):597–604. [PMC free article] [PubMed]
  • McBurney MW, Jones-Villeneuve EM, Edwards MK, Anderson PJ. Control of muscle and neuronal differentiation in a cultured embryonal carcinoma cell line. Nature. 1982 Sep 9;299(5879):165–167. [PubMed]
  • Melton DA, Krieg PA, Rebagliati MR, Maniatis T, Zinn K, Green MR. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. [PMC free article] [PubMed]
  • Molony L, Burridge K. Molecular shape and self-association of vinculin and metavinculin. J Cell Biochem. 1985;29(1):31–36. [PubMed]
  • Nagafuchi A, Takeichi M. Cell binding function of E-cadherin is regulated by the cytoplasmic domain. EMBO J. 1988 Dec 1;7(12):3679–3684. [PubMed]
  • Nagafuchi A, Takeichi M, Tsukita S. The 102 kd cadherin-associated protein: similarity to vinculin and posttranscriptional regulation of expression. Cell. 1991 May 31;65(5):849–857. [PubMed]
  • Nagafuchi A, Takeichi M. Transmembrane control of cadherin-mediated cell adhesion: a 94 kDa protein functionally associated with a specific region of the cytoplasmic domain of E-cadherin. Cell Regul. 1989 Nov;1(1):37–44. [PMC free article] [PubMed]
  • Nelson WJ, Veshnock PJ. Dynamics of membrane-skeleton (fodrin) organization during development of polarity in Madin-Darby canine kidney epithelial cells. J Cell Biol. 1986 Nov;103(5):1751–1765. [PMC free article] [PubMed]
  • Otey CA, Pavalko FM, Burridge K. An interaction between alpha-actinin and the beta 1 integrin subunit in vitro. J Cell Biol. 1990 Aug;111(2):721–729. [PMC free article] [PubMed]
  • Otto JJ. Vinculin. Cell Motil Cytoskeleton. 1990;16(1):1–6. [PubMed]
  • Ozawa M, Baribault H, Kemler R. The cytoplasmic domain of the cell adhesion molecule uvomorulin associates with three independent proteins structurally related in different species. EMBO J. 1989 Jun;8(6):1711–1717. [PubMed]
  • Ozawa M, Ringwald M, Kemler R. Uvomorulin-catenin complex formation is regulated by a specific domain in the cytoplasmic region of the cell adhesion molecule. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4246–4250. [PubMed]
  • Peyriéras N, Louvard D, Jacob F. Characterization of antigens recognized by monoclonal and polyclonal antibodies directed against uvomorulin. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8067–8071. [PubMed]
  • Pfeiffer DC, Vogl AW. Evidence that vinculin is co-distributed with actin bundles in ectoplasmic ("junctional") specializations of mammalian Sertoli cells. Anat Rec. 1991 Sep;231(1):89–100. [PubMed]
  • Price GJ, Jones P, Davison MD, Patel B, Bendori R, Geiger B, Critchley DR. Primary sequence and domain structure of chicken vinculin. Biochem J. 1989 Apr 15;259(2):453–461. [PubMed]
  • Reszka AA, Hayashi Y, Horwitz AF. Identification of amino acid sequences in the integrin beta 1 cytoplasmic domain implicated in cytoskeletal association. J Cell Biol. 1992 Jun;117(6):1321–1330. [PMC free article] [PubMed]
  • Ryseck RP, MacDonald-Bravo H, Zerial M, Bravo R. Coordinate induction of fibronectin, fibronectin receptor, tropomyosin, and actin genes in serum-stimulated fibroblasts. Exp Cell Res. 1989 Feb;180(2):537–545. [PubMed]
  • Shirinsky VP, Birukov KG, Koteliansky VE, Glukhova MA, Spanidis E, Rogers JD, Campbell JH, Campbell GR. Density-related expression of caldesmon and vinculin in cultured rabbit aortic smooth muscle cells. Exp Cell Res. 1991 Jun;194(2):186–189. [PubMed]
  • Tsukita S, Tsukita S, Nagafuchi A, Yonemura S. Molecular linkage between cadherins and actin filaments in cell-cell adherens junctions. Curr Opin Cell Biol. 1992 Oct;4(5):834–839. [PubMed]
  • Tsukita S, Itoh M, Tsukita S. A new 400-kD protein from isolated adherens junctions: its localization at the undercoat of adherens junctions and at microfilament bundles such as stress fibers and circumferential bundles. J Cell Biol. 1989 Dec;109(6 Pt 1):2905–2915. [PMC free article] [PubMed]
  • Turner CE, Burridge K. Transmembrane molecular assemblies in cell-extracellular matrix interactions. Curr Opin Cell Biol. 1991 Oct;3(5):849–853. [PubMed]
  • Turner CE, Glenney JR, Jr, Burridge K. Paxillin: a new vinculin-binding protein present in focal adhesions. J Cell Biol. 1990 Sep;111(3):1059–1068. [PMC free article] [PubMed]
  • Ungar F, Geiger B, Ben-Ze'ev A. Cell contact- and shape-dependent regulation of vinculin synthesis in cultured fibroblasts. Nature. 319(6056):787–791. [PubMed]
  • Vestweber D, Kemler R. Some structural and functional aspects of the cell adhesion molecule uvomorulin. Cell Differ. 1984 Dec;15(2-4):269–273. [PubMed]
  • Weller PA, Ogryzko EP, Corben EB, Zhidkova NI, Patel B, Price GJ, Spurr NK, Koteliansky VE, Critchley DR. Complete sequence of human vinculin and assignment of the gene to chromosome 10. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5667–5671. [PubMed]
  • Westmeyer A, Ruhnau K, Wegner A, Jockusch BM. Antibody mapping of functional domains in vinculin. EMBO J. 1990 Jul;9(7):2071–2078. [PubMed]
  • Wood CM, Moses RE. Ethyl methane sulfonate- and bleomycin-generated deletion mutations at HPRT locus in xeroderma pigmentosum complementation group D fibroblasts. Somat Cell Mol Genet. 1989 Jul;15(4):345–357. [PubMed]

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