Home LiteratureArticle Details
PMID: 1671296 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The transforming potential of the c-erbB-2 protein is regulated by its autophosphorylation at the carboxyl-terminal domain.

Molecular and cellular biology ·Vol. 11 ·No. 2 ·1991-02-00 ·Pages 833-42

Akiyama T, Matsuda S, Namba Y, Saito T, Toyoshima K, Yamamoto T

Abstract

The mutant c-erbB-2 protein with Glu instead of Val-659 exhibited transforming activity in NIH 3T3 cells. This protein showed enhanced tyrosine kinase activity in vitro and enhanced autophosphorylation at Tyr-1248 located proximal to the carboxyl terminus. Enhanced tyrosine phosphorylation of several cellular proteins was detected in cells expressing the Glu-659 c-erbB-2 protein. Introduction of an additional mutation at the ATP-binding site (Lys-753 to Met) of this protein resulted in abolition of its transforming ability. These data indicate that the transforming potential of c-erbB-2 is closely correlated with elevated tyrosine kinase activity of the gene product. To investigate the role of autophosphorylation in cell transformation, we introduced an additional mutation at the autophosphorylation site of the Glu-659 c-erbB-2 protein (Tyr-1248 to Phe). This mutant protein exhibited lower tyrosine kinase activity and lower transforming activity. On the other hand, when the carboxyl-terminal 230 amino acid residues were deleted from the c-erbB-2 protein, the tyrosine kinase activity and cell-transforming activity of the protein were enhanced. Thus, the carboxyl-terminal domain, which contains the major autophosphorylation site, Tyr-1248, may regulate cellular transformation negatively and autophosphorylation may eliminate this negative regulation.

Related Genes
c-erbB-2
MeSH Terms
Animals Base Sequence Cell Division Cell Line Cell Transformation, Neoplastic Kinetics Mice Molecular Sequence Data Mutagenesis, Site-Directed Oligonucleotide Probes Phosphorylation Protein-Tyrosine Kinases/metabolism Proto-Oncogene Proteins/genetics,isolation & purification,metabolism Proto-Oncogenes Receptor, ErbB-2 Transfection
Chemicals
Oligonucleotide Probes Proto-Oncogene Proteins Protein-Tyrosine Kinases Receptor, ErbB-2
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Akiyama T
Department of Oncogene Research, Osaka University, Japan.
Matsuda S
Namba Y
Saito T
Toyoshima K
Yamamoto T
References (43)
43 references, click to expand
  1. Identification of multiple novel polypeptide substrates of the v-src, v-yes, v-fps, v-ros, and v-erb-B oncogenic tyrosine protein kinases utilizing antisera against phosphotyrosine.
    Oncogene. 1988 Apr;2(4):305-15 PMID: 2452398
  2. Purification of the catalytically active phosphorylated form of insulin receptor kinase by affinity chromatography with O-phosphotyrosyl-binding antibodies.
    Arch Biochem Biophys. 1985 Oct;242(1):176-86 PMID: 2413806
  3. Autophosphorylation sites on the epidermal growth factor receptor.
    Nature. 1984 Oct 4-10;311(5985):483-5 PMID: 6090945
  4. Subversion of growth regulatory pathways in malignant transformation.
    Biochim Biophys Acta. 1987 Nov 25;907(3):219-44 PMID: 3314997
  5. The neu oncogene encodes an epidermal growth factor receptor-related protein.
    Nature. 1986 Jan 16-22;319(6050):226-30 PMID: 3945311
  6. Phosphorylation of GAP and GAP-associated proteins by transforming and mitogenic tyrosine kinases.
    Nature. 1990 Jan 25;343(6256):377-81 PMID: 1689011
  7. Epidermal growth factor stimulates the phosphorylation of synthetic tyrosine-containing peptides by A431 cell membranes.
    Proc Natl Acad Sci U S A. 1982 Mar;79(5):1443-7 PMID: 6175968
  8. The role of autophosphorylation in modulation of erbB-2 transforming function.
    New Biol. 1990 Feb;2(2):187-95 PMID: 1982072
  9. Identification of autophosphorylation sites of HER2/neu.
    Cell Growth Differ. 1990 Jan;1(1):3-7 PMID: 1706616
  10. Direct interaction of a ligand for the erbB2 oncogene product with the EGF receptor and p185erbB2.
    Science. 1990 Sep 28;249(4976):1552-5 PMID: 2218496
  11. C-erbB-2 gene product, a membrane protein commonly expressed on human fetal epithelial cells.
    Lab Invest. 1989 Jul;61(1):93-7 PMID: 2473278
  12. Phospholipase C-gamma is a substrate for the PDGF and EGF receptor protein-tyrosine kinases in vivo and in vitro.
    Cell. 1989 Jun 30;57(7):1109-22 PMID: 2472219
  13. EGF induces tyrosine phosphorylation of phospholipase C-II: a potential mechanism for EGF receptor signaling.
    Cell. 1989 Jun 30;57(7):1101-7 PMID: 2472218
  14. Analysis of mammalian fibroblast transformation by normal and mutated human EGF receptors.
    Oncogene. 1989 Mar;4(3):273-83 PMID: 2784850
  15. Oncogenic activation of p185neu stimulates tyrosine phosphorylation in vivo.
    Mol Cell Biol. 1988 Sep;8(9):3969-73 PMID: 2464744
  16. Epidermal growth factor stimulates tyrosine phosphorylation of phospholipase C-II independently of receptor internalization and extracellular calcium.
    Proc Natl Acad Sci U S A. 1989 Mar;86(5):1568-72 PMID: 2466293
  17. Biological activities of EGF-receptor mutants with individually altered autophosphorylation sites.
    EMBO J. 1988 Oct;7(10):3045-52 PMID: 3263271
  18. Signal transduction from membrane to cytoplasm: growth factors and membrane-bound oncogene products increase Raf-1 phosphorylation and associated protein kinase activity.
    Proc Natl Acad Sci U S A. 1988 Dec;85(23):8855-9 PMID: 3057494
  19. A Mr = 190,000 glycoprotein phosphorylated on tyrosine residues in epidermal growth factor stimulated KB cells is the product of the c-erbB-2 gene.
    Biochem Biophys Res Commun. 1987 Apr 29;144(2):699-704 PMID: 2437918
  20. erbB-2 is a potent oncogene when overexpressed in NIH/3T3 cells.
    Science. 1987 Jul 10;237(4811):178-82 PMID: 2885917
  21. Requirement for intrinsic protein tyrosine kinase in the immediate and late actions of the EGF receptor.
    Nature. 1987 Aug 27-Sep 2;328(6133):820-3 PMID: 3498122
  22. Alteration of epidermal growth factor receptor activity by mutation of its primary carboxyl-terminal site of tyrosine self-phosphorylation.
    J Biol Chem. 1988 Mar 15;263(8):3610-7 PMID: 3346210
  23. Growth factor control of epidermal growth factor receptor kinase activity via an intramolecular mechanism.
    J Biol Chem. 1988 Feb 15;263(5):2230-7 PMID: 3257489
  24. Stage- and tissue-specific expression of the neu oncogene in rat development.
    Proc Natl Acad Sci U S A. 1987 Dec;84(23):8498-501 PMID: 3317411
  25. Nonmyristoylated p60v-src fails to phosphorylate proteins of 115-120 kDa in chicken embryo fibroblasts.
    Proc Natl Acad Sci U S A. 1988 Apr;85(8):2608-12 PMID: 2451829
  26. Human insulin receptors mutated at the ATP-binding site lack protein tyrosine kinase activity and fail to mediate postreceptor effects of insulin.
    J Biol Chem. 1987 Feb 5;262(4):1842-7 PMID: 3100537
  27. Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene.
    Science. 1987 Jan 9;235(4785):177-82 PMID: 3798106
  28. p185, a product of the neu proto-oncogene, is a receptorlike protein associated with tyrosine kinase activity.
    Mol Cell Biol. 1986 May;6(5):1729-40 PMID: 2878363
  29. Structural requirements for the transmembrane activation of the insulin receptor kinase.
    J Biol Chem. 1986 Nov 15;261(32):15281-7 PMID: 3021769
  30. Substrate specificities of tyrosine-specific protein kinases toward cytoskeletal proteins in vitro.
    J Biol Chem. 1986 Nov 5;261(31):14797-803 PMID: 3771552
  31. Tumor promoter and epidermal growth factor stimulate phosphorylation of the c-erbB-2 gene product in MKN-7 human adenocarcinoma cells.
    Mol Cell Biol. 1988 Mar;8(3):1019-26 PMID: 2897079
  32. Intrapeptide autophosphorylation of the epidermal growth factor receptor: regulation of kinase catalytic function by receptor dimerization.
    Biochemistry. 1985 Jul 2;24(14):3795-802 PMID: 2994718
  33. Protein-tyrosine kinases.
    Annu Rev Biochem. 1985;54:897-930 PMID: 2992362
  34. Self-phosphorylation enhances the protein-tyrosine kinase activity of the epidermal growth factor receptor.
    J Biol Chem. 1985 Nov 25;260(27):14642-7 PMID: 2997217
  35. Autophosphorylation and protein kinase C phosphorylation of the epidermal growth factor receptor. Effect on tyrosine kinase activity and ligand binding affinity.
    J Biol Chem. 1985 Nov 25;260(27):14538-46 PMID: 2997213
  36. The product of the human c-erbB-2 gene: a 185-kilodalton glycoprotein with tyrosine kinase activity.
    Science. 1986 Jun 27;232(4758):1644-6 PMID: 3012781
  37. Amplification of c-erbB-2 oncogene in human adenocarcinomas in vivo.
    Lancet. 1986 Apr 5;1(8484):765-7 PMID: 2870269
  38. The rapid generation of oligonucleotide-directed mutations at high frequency using phosphorothioate-modified DNA.
    Nucleic Acids Res. 1985 Dec 20;13(24):8765-85 PMID: 3001650
  39. Oncogenic activation of the neu-encoded receptor protein by point mutation and deletion.
    EMBO J. 1988 Jul;7(7):2043-52 PMID: 2901345
  40. Tyrosine kinase receptor with extensive homology to EGF receptor shares chromosomal location with neu oncogene.
    Science. 1985 Dec 6;230(4730):1132-9 PMID: 2999974
  41. A v-erbB-related protooncogene, c-erbB-2, is distinct from the c-erbB-1/epidermal growth factor-receptor gene and is amplified in a human salivary gland adenocarcinoma.
    Proc Natl Acad Sci U S A. 1985 Oct;82(19):6497-501 PMID: 2995967
  42. Amplification of a novel v-erbB-related gene in a human mammary carcinoma.
    Science. 1985 Sep 6;229(4717):974-6 PMID: 2992089
  43. Genetic alterations of the c-erbB-2 oncogene occur frequently in tubular adenocarcinoma of the stomach and are often accompanied by amplification of the v-erbA homologue.
    Oncogene. 1988 Mar;2(3):283-7 PMID: 3281095
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.11.2.833-842.1991
Published
1991-02-00
Pages
833-42
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC359735
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.11.2.833-842.1991 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