Home LiteratureArticle Details
PMID: 3117046 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Isolation and characterization of a vinculin cDNA from chick-embryo fibroblasts.

The Biochemical journal ·Vol. 245 ·No. 2 ·1987-07-15 ·Pages 595-603

Price GJ, Jones P, Davison MD, Patel B, Eperon IC, Critchley DR

Abstract

A chick-embryo fibroblast lambda gt11 cDNA library was screened with affinity-purified antibodies to chick gizzard vinculin. One recombinant was purified to homogeneity and the fusion protein expressed in Escherichia coli strain C600. The fusion protein was unstable, but polypeptides that reacted with vinculin antibodies, but not non-immune immunoglobulin, were detected by Western blotting. The recombinant contained a single EcoRI fragment of 2891 bp with a single open reading frame. The deduced protein sequence could be aligned with that of six CNBr-cleavage peptides and two tryptic peptides derived from chicken gizzard vinculin. AUG-247 has tentatively been identified as the initiation codon, as it is contained within the consensus sequence for initiation sites of higher eukaryotes. The cDNA lacks 3' sequence and encodes 74% of the vinculin sequence, presuming the molecular mass of vinculin to be 130,000 Da. Analysis of the deduced sequence showed no homologies with other protein sequences, but it does display a triple internal repeat of 112 amino acid residues covering residues 259-589. The sequences surrounding the seven tyrosine residues in the available sequence were aligned with the tyrosine autophosphorylation consensus sequence found in protein tyrosine kinases. Tyr-822 showed a good match to this consensus, and may represent one of the two major sites of tyrosine phosphorylation by pp60v-sre. Northern blots showed that the 2.89 kb vinculin cDNA hybridized to one size of mRNA (approx. 7 kb) in chick-embryo fibroblasts, chick smooth muscle and chick skeletal muscle. Southern blots revealed multiple hybridizing bands in genomic DNA.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Binding Sites Chick Embryo DNA/isolation & purification Fibroblasts/analysis Immunoelectrophoresis Muscle Proteins/genetics Nucleic Acid Hybridization RNA, Messenger/genetics Tyrosine/analysis Vinculin
Chemicals
Muscle Proteins RNA, Messenger Vinculin Tyrosine DNA
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Price G J
Department of Biochemistry, University of Leicester, U.K.
Jones P
Davison M D
Patel B
Eperon I C
Critchley D R
References (59)
59 references, click to expand
  1. The chromatographic determination of cystine and cysteine residues in proteins as s-beta-(4-pyridylethyl)cysteine.
    J Biol Chem. 1970 Aug 10;245(15):3868-71 PMID: 5492953
  2. The cytoskeletal protein vinculin is acylated by myristic acid.
    FEBS Lett. 1987 Mar 23;213(2):428-32 PMID: 3030822
  3. Amino acid sequence of beta-galactosidase. XI. Peptide ordering procedures and the complete sequence.
    J Biol Chem. 1978 Aug 10;253(15):5521-5 PMID: 97298
  4. Alpha-actinin: immunofluorescent localization of a muscle structural protein in nonmuscle cells.
    Cell. 1975 Nov;6(3):289-98 PMID: 802682
  5. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4 PMID: 388439
  6. 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):4127-31 PMID: 6776523
  7. Vinculin: a cytoskeletal target of the transforming protein of Rous sarcoma virus.
    Cell. 1981 Apr;24(1):165-74 PMID: 6263485
  8. An interactive graphics program for comparing and aligning nucleic acid and amino acid sequences.
    Nucleic Acids Res. 1982 May 11;10(9):2951-61 PMID: 7099970
  9. Relationships between microfilaments, cell-substratum adhesion, and fibronectin.
    Cold Spring Harb Symp Quant Biol. 1982;46 Pt 2:659-70 PMID: 7049533
  10. Mechanism of transformation by Rous sarcoma virus: events within adhesion plaques.
    Cold Spring Harb Symp Quant Biol. 1982;46 Pt 2:953-65 PMID: 6286226
  11. Co-existence of vinculin and a vinculin-like protein of higher molecular weight in smooth muscle.
    J Biol Chem. 1982 Sep 25;257(18):11024-31 PMID: 6809764
  12. Automation of the computer handling of gel reading data produced by the shotgun method of DNA sequencing.
    Nucleic Acids Res. 1982 Aug 11;10(15):4731-51 PMID: 7133997
  13. Microheterogeneity of avian and mammalian vinculin distinctive subcellular distribution of different isovinculins.
    J Mol Biol. 1982 Aug 25;159(4):685-701 PMID: 6815331
  14. Meta-vinculin--a vinculin-related protein with solubility properties of a membrane protein.
    Nature. 1982 Dec 9;300(5892):533-5 PMID: 6815540
  15. Statistical characterization of nucleic acid sequence functional domains.
    Nucleic Acids Res. 1983 Apr 11;11(7):2205-20 PMID: 6835847
  16. A new protein of adhesion plaques and ruffling membranes.
    J Cell Biol. 1983 Aug;97(2):359-67 PMID: 6684120
  17. Vinculin phosphorylation by the src kinase. Interaction of vinculin with phospholipid vesicles.
    J Biol Chem. 1983 Dec 10;258(23):14626-31 PMID: 6417142
  18. Gene product of v-fgr onc: hybrid protein containing a portion of actin and a tyrosine-specific protein kinase.
    Science. 1984 Jan 6;223(4631):63-6 PMID: 6318314
  19. Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs.
    Nucleic Acids Res. 1984 Jan 25;12(2):857-72 PMID: 6694911
  20. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis.
    Gene. 1983 Dec;26(1):101-6 PMID: 6323249
  21. Properties of smooth muscle vinculin.
    J Biol Chem. 1984 Mar 25;259(6):3916-24 PMID: 6323447
  22. Sequence variability in the retinal-attachment domain of mammalian rhodopsins.
    Biochem J. 1984 Feb 1;217(3):605-13 PMID: 6370231
  23. Vinculin phosphorylation in response to calcium and phorbol esters in intact cells.
    J Biol Chem. 1984 Apr 25;259(8):5264-70 PMID: 6325428
  24. The role of protein kinase C in cell surface signal transduction and tumour promotion.
    Nature. 1984 Apr 19-25;308(5961):693-8 PMID: 6232463
  25. An interaction between vinculin and talin.
    Nature. 1984 Apr 19-25;308(5961):744-6 PMID: 6425696
  26. Cell attachment activity of fibronectin can be duplicated by small synthetic fragments of the molecule.
    Nature. 1984 May 3-9;309(5963):30-3 PMID: 6325925
  27. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum.
    Anal Biochem. 1984 Feb;137(1):266-7 PMID: 6329026
  28. Phosphorylation sites in enolase and lactate dehydrogenase utilized by tyrosine protein kinases in vivo and in vitro.
    J Biol Chem. 1984 Jun 25;259(12):7835-41 PMID: 6330085
  29. Isolation and some properties of macrophage alpha-actinin: evidence that it is not an actin gelling protein.
    Biochemistry. 1984 Oct 9;23(21):5081-6 PMID: 6498177
  30. Amino terminal myristylation of the protein kinase p60src, a retroviral transforming protein.
    Science. 1985 Jan 25;227(4685):427-9 PMID: 3917576
  31. Identification and isolation of a 140 kd cell surface glycoprotein with properties expected of a fibronectin receptor.
    Cell. 1985 Jan;40(1):191-8 PMID: 3155652
  32. Platelet-derived growth factor-induced alterations in vinculin and actin distribution in BALB/c-3T3 cells.
    J Cell Biol. 1985 Apr;100(4):1031-40 PMID: 3920222
  33. Increased phosphorylation of tyrosine in vinculin does not occur upon transformation by some avian sarcoma viruses.
    Mol Cell Biol. 1985 Jan;5(1):263-7 PMID: 2580230
  34. Distribution of the cell substratum attachment (CSAT) antigen on myogenic and fibroblastic cells in culture.
    J Cell Biol. 1985 May;100(5):1528-39 PMID: 3921554
  35. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  36. The use of synthetic oligonucleotides with universal templates for rapid DNA sequencing: results with staphylococcal replicon pC221.
    EMBO J. 1985 Feb;4(2):561-8 PMID: 3860383
  37. Protein-tyrosine kinases.
    Annu Rev Biochem. 1985;54:897-930 PMID: 2992362
  38. Electron microscopy of rotary shadowed vinculin and vinculin complexes.
    J Mol Biol. 1985 Aug 5;184(3):543-5 PMID: 3930754
  39. Posttranslational processing of p21 ras proteins involves palmitylation of the C-terminal tetrapeptide containing cysteine-186.
    J Virol. 1985 Nov;56(2):607-12 PMID: 2997480
  40. A human oncogene formed by the fusion of truncated tropomyosin and protein tyrosine kinase sequences.
    Nature. 1986 Feb 27-Mar 5;319(6056):743-8 PMID: 2869410
  41. Human oestrogen receptor cDNA: sequence, expression and homology to v-erb-A.
    Nature. 1986 Mar 13-19;320(6058):134-9 PMID: 3754034
  42. Reorganization of alpha-actinin and vinculin induced by a phorbol ester in living cells.
    J Cell Biol. 1986 Apr;102(4):1430-8 PMID: 3082892
  43. Cell biology. Finding the missing links.
    Nature. 1986 Apr 10-16;320(6062):484-5 PMID: 3960130
  44. Interaction of plasma membrane fibronectin receptor with talin--a transmembrane linkage.
    Nature. 1986 Apr 10-16;320(6062):531-3 PMID: 2938015
  45. Interaction of the cytoskeletal component vinculin with bilayer structures analyzed with a photoactivatable phospholipid.
    J Biol Chem. 1986 May 25;261(15):6912-8 PMID: 3084492
  46. Stress fibers in the splenic sinus endothelium in situ: molecular structure, relationship to the extracellular matrix, and contractility.
    J Cell Biol. 1986 May;102(5):1738-47 PMID: 3084499
  47. Identification of actin-, alpha-actinin-, and vinculin-containing plaques at the lateral membrane of epithelial cells.
    J Cell Biol. 1986 May;102(5):1843-52 PMID: 3084501
  48. The use of Rous sarcoma virus transformation mutants with differing tyrosine kinase activities to study the relationships between vinculin phosphorylation, pp60v-src location and adhesion plaque integrity.
    Exp Cell Res. 1986 Jul;165(1):216-28 PMID: 3011478
  49. Length polymorphism in the pro alpha 2(I) collagen gene: an alternative explanation in a case of Marfan syndrome.
    Hum Genet. 1986 May;73(1):91-2 PMID: 3011649
  50. The cDNA sequence for the protein-tyrosine kinase substrate p36 (calpactin I heavy chain) reveals a multidomain protein with internal repeats.
    Cell. 1986 Jul 18;46(2):201-12 PMID: 3013423
  51. Structure of integrin, a glycoprotein involved in the transmembrane linkage between fibronectin and actin.
    Cell. 1986 Jul 18;46(2):271-82 PMID: 3487386
  52. Talin is phosphorylated on tyrosine in chicken embryo fibroblasts transformed by Rous sarcoma virus.
    Proc Natl Acad Sci U S A. 1986 Aug;83(15):5507-11 PMID: 2426700
  53. The complete primary structure of protein kinase C--the major phorbol ester receptor.
    Science. 1986 Aug 22;233(4766):853-9 PMID: 3755547
  54. Phosphorylation of the fibronectin receptor complex in cells transformed by oncogenes that encode tyrosine kinases.
    Proc Natl Acad Sci U S A. 1986 Sep;83(17):6470-4 PMID: 3018734
  55. The mRNA and RNA-copy pseudogenes encoding TM30nm, a human cytoskeletal tropomyosin.
    Nucleic Acids Res. 1986 Nov 11;14(21):8413-26 PMID: 3024106
  56. Comparison of the relative importance of tyrosine-specific vinculin phosphorylation and the loss of surface-associated fibronectin in the morphology of cells transformed by Rous sarcoma virus.
    J Cell Sci. 1986 Jun;82:129-42 PMID: 3025229
  57. The cytoskeletal protein vinculin contains transformation-sensitive, covalently bound lipid.
    Science. 1987 Jan 23;235(4787):476-9 PMID: 3099391
  58. Isolation and characterization of a cDNA encoding a chick alpha-actinin.
    J Biol Chem. 1987 Feb 25;262(6):2558-61 PMID: 3818609
  59. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2450595
Published
1987-07-15
Pages
595-603
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1148163
Subset
IM
Databases
GENBANK
Y00312
External Links
PubMed source DOI: 10.1042/bj2450595 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: [email protected]

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