Home LiteratureArticle Details
PMID: 8622698 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Truncated mammalian Notch1 activates CBF1/RBPJk-repressed genes by a mechanism resembling that of Epstein-Barr virus EBNA2.

Molecular and cellular biology ·Vol. 16 ·No. 3 ·1996-03-00 ·Pages 952-9

Hsieh JJ, Henkel T, Salmon P, Robey E, Peterson MG, Hayward SD

Abstract

The Notch/Lin-12/Glp-1 receptor family participates in cell-cell signaling events that influence cell fate decisions. Although several Notch homologs and receptor ligands have been identified, the nuclear events involved in this pathway remain incompletely understood. A truncated form of Notch, consisting only of the intracellular domain (NotchIC), localizes to the nucleus and functions as an activated receptor. Using both an in vitro binding assay and a cotransfection assay based on the two-hybrid principle, we show that mammalian NotchIC interacts with the transcriptional repressor CBF1, which is the human homolog of Drosophila Suppressor of Hairless. Cotransfection assays using segments of mouse NotchIC and CBF1 demonstrated that the N-terminal 114-amino-acid region of mouse NotchIC contains the CBF1 interactive domain and that the cdc10/ankyrin repeats are not essential for this interaction. This result was confirmed in immunoprecipation assays in which the N-terminal 114-amino-acid segment of NotchIC, but not the ankyrin repeat region, coprecipitated with CBF1. Mouse NotchIC itself is targeted to the transcriptional repression domain (aa179 to 361) of CBF1. Furthermore, transfection assays in which mouse NotchIC was targeted through Gal4-CBF1 or through endogenous cellular CBF1 indicated that NotchIC transactivates gene expression via CBF1 tethering to DNA. Transactivation by NotchIC occurs partially through abolition of CBF1-mediated repession. This same mechanism is used by Epstein-Barr virus EBNA2. Thus, mimicry of Notch signal transduction is involved in Epstein-Barr virus-driven immortalization.

MeSH Terms
Amino Acid Sequence Animals Antigens, Viral/genetics,metabolism Base Sequence Basic Helix-Loop-Helix Leucine Zipper Transcription Factors DNA-Binding Proteins/genetics,metabolism Drosophila Proteins Drosophila melanogaster Epstein-Barr Virus Nuclear Antigens Fungal Proteins/genetics Gene Expression Regulation Gene Transfer Techniques HeLa Cells Humans Membrane Proteins/genetics,metabolism Molecular Sequence Data Receptors, Notch Repressor Proteins/genetics,metabolism Saccharomyces cerevisiae Proteins Sequence Alignment Signal Transduction
Chemicals
Antigens, Viral Basic Helix-Loop-Helix Leucine Zipper Transcription Factors CBF1 protein, S cerevisiae DNA-Binding Proteins Drosophila Proteins Epstein-Barr Virus Nuclear Antigens Fungal Proteins Membrane Proteins N protein, Drosophila Receptors, Notch Repressor Proteins Saccharomyces cerevisiae Proteins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Hsieh J J
Department of Pharmacology and Molecular Sciences, The Johns Hopkins University of Medicine, Baltimore, Maryland 21205, USA.
Henkel T
Salmon P
Robey E
Peterson M G
Hayward S D
References (63)
63 references, click to expand
  1. Suppressor of Hairless is required for signal reception during lateral inhibition in the Drosophila pupal notum.
    Development. 1995 Jun;121(6):1875-84 PMID: 7601001
  2. Expression of a Delta homologue in prospective neurons in the chick.
    Nature. 1995 Jun 29;375(6534):787-90 PMID: 7596411
  3. lag-2 may encode a signaling ligand for the GLP-1 and LIN-12 receptors of C. elegans.
    Development. 1994 Oct;120(10):2913-24 PMID: 7607081
  4. Constitutive expression of a truncated INT3 gene in mouse mammary epithelium impairs differentiation and functional development.
    Cell Growth Differ. 1995 May;6(5):563-77 PMID: 7544153
  5. Signalling downstream of activated mammalian Notch.
    Nature. 1995 Sep 28;377(6547):355-8 PMID: 7566092
  6. The suppressor of hairless protein participates in notch receptor signaling.
    Cell. 1994 Oct 21;79(2):273-82 PMID: 7954795
  7. Nucleotide sequence from the neurogenic locus notch implies a gene product that shares homology with proteins containing EGF-like repeats.
    Cell. 1985 Dec;43(3 Pt 2):567-81 PMID: 3935325
  8. Mutations altering the structure of epidermal growth factor-like coding sequences at the Drosophila Notch locus.
    Cell. 1987 Nov 20;51(4):539-48 PMID: 3119223
  9. The Caenorhabditis elegans lin-12 gene encodes a transmembrane protein with overall similarity to Drosophila Notch.
    Nature. 1988 Oct 6;335(6190):547-50 PMID: 3419531
  10. Genetic analysis of immortalizing functions of Epstein-Barr virus in human B lymphocytes.
    Nature. 1989 Aug 3;340(6232):393-7 PMID: 2547164
  11. glp-1 and lin-12, genes implicated in distinct cell-cell interactions in C. elegans, encode similar transmembrane proteins.
    Cell. 1989 Aug 11;58(3):553-63 PMID: 2758466
  12. Transcript analysis of glp-1 and lin-12, homologous genes required for cell interactions during development of C. elegans.
    Cell. 1989 Aug 11;58(3):565-71 PMID: 2758467
  13. A protein binding to the J kappa recombination sequence of immunoglobulin genes contains a sequence related to the integrase motif.
    Nature. 1989 Dec 21-28;342(6252):934-7 PMID: 2556644
  14. Epstein-Barr virus nuclear protein 2 is a key determinant of lymphocyte transformation.
    Proc Natl Acad Sci U S A. 1989 Dec;86(23):9558-62 PMID: 2556717
  15. Molecular interactions between the protein products of the neurogenic loci Notch and Delta, two EGF-homologous genes in Drosophila.
    Cell. 1990 May 4;61(3):523-34 PMID: 2185893
  16. Isolation and characterization of the Drosophila gene encoding the TATA box binding protein, TFIID.
    Cell. 1990 Jun 29;61(7):1179-86 PMID: 2194666
  17. Xotch, the Xenopus homolog of Drosophila notch.
    Science. 1990 Sep 21;249(4975):1438-41 PMID: 2402639
  18. deltex, a locus interacting with the neurogenic genes, Notch, Delta and mastermind in Drosophila melanogaster.
    Genetics. 1990 Nov;126(3):665-77 PMID: 2123462
  19. Role for the Epstein-Barr virus nuclear antigen 2 in viral promoter switching during initial stages of infection.
    Proc Natl Acad Sci U S A. 1991 May 1;88(9):3942-6 PMID: 1850841
  20. TAN-1, the human homolog of the Drosophila notch gene, is broken by chromosomal translocations in T lymphoblastic neoplasms.
    Cell. 1991 Aug 23;66(4):649-61 PMID: 1831692
  21. A role for the Drosophila neurogenic genes in mesoderm differentiation.
    Cell. 1991 Oct 18;67(2):311-23 PMID: 1913825
  22. Specific EGF repeats of Notch mediate interactions with Delta and Serrate: implications for Notch as a multifunctional receptor.
    Cell. 1991 Nov 15;67(4):687-99 PMID: 1657403
  23. A homolog of Drosophila Notch expressed during mammalian development.
    Development. 1991 Sep;113(1):199-205 PMID: 1764995
  24. Expression of an activated Notch-related int-3 transgene interferes with cell differentiation and induces neoplastic transformation in mammary and salivary glands.
    Genes Dev. 1992 Mar;6(3):345-55 PMID: 1372276
  25. Mouse mammary tumor gene int-3: a member of the notch gene family transforms mammary epithelial cells.
    J Virol. 1992 Apr;66(4):2594-9 PMID: 1312643
  26. A genetic analysis of deltex and its interaction with the Notch locus in Drosophila melanogaster.
    Genetics. 1992 May;131(1):99-112 PMID: 1592246
  27. The Drosophila homolog of the immunoglobulin recombination signal-binding protein regulates peripheral nervous system development.
    Cell. 1992 Jun 26;69(7):1191-7 PMID: 1617729
  28. Suppressor of Hairless, the Drosophila homolog of the mouse recombination signal-binding protein gene, controls sensory organ cell fates.
    Cell. 1992 Jun 26;69(7):1199-212 PMID: 1617730
  29. Molecular basis of loss-of-function mutations in the glp-1 gene of Caenorhabditis elegans.
    Mol Biol Cell. 1992 Nov;3(11):1199-213 PMID: 1457827
  30. Cloning, analysis, and chromosomal localization of Notch-1, a mouse homolog of Drosophila Notch.
    Genomics. 1993 Feb;15(2):259-64 PMID: 8449489
  31. The function of the neurogenic genes during epithelial development in the Drosophila embryo.
    Development. 1992 Dec;116(4):1203-20 PMID: 1295737
  32. Human homologs of a Drosophila Enhancer of split gene product define a novel family of nuclear proteins.
    Nat Genet. 1992 Oct;2(2):119-27 PMID: 1303260
  33. EBNA-2 of herpesvirus papio diverges significantly from the type A and type B EBNA-2 proteins of Epstein-Barr virus but retains an efficient transactivation domain with a conserved hydrophobic motif.
    J Virol. 1993 Jun;67(6):2990-3003 PMID: 8388484
  34. Expression of an extracellular deletion of Xotch diverts cell fate in Xenopus embryos.
    Cell. 1993 May 21;73(4):659-71 PMID: 8500162
  35. Specific truncations of Drosophila Notch define dominant activated and dominant negative forms of the receptor.
    Cell. 1993 Jul 30;74(2):319-29 PMID: 8343959
  36. Intrinsic activity of the Lin-12 and Notch intracellular domains in vivo.
    Cell. 1993 Jul 30;74(2):331-45 PMID: 8343960
  37. Antineurogenic phenotypes induced by truncated Notch proteins indicate a role in signal transduction and may point to a novel function for Notch in nuclei.
    Genes Dev. 1993 Oct;7(10):1949-65 PMID: 8406001
  38. Human Jk recombination signal binding protein gene (IGKJRB): comparison with its mouse homologue.
    Genomics. 1993 Aug;17(2):306-15 PMID: 8406481
  39. An activated Notch receptor blocks cell-fate commitment in the developing Drosophila eye.
    Nature. 1993 Oct 7;365(6446):555-7 PMID: 8413612
  40. The Epstein-Barr virus immortalizing protein EBNA-2 is targeted to DNA by a cellular enhancer-binding protein.
    Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9237-41 PMID: 8415684
  41. A zebrafish homologue of the Drosophila neurogenic gene Notch and its pattern of transcription during early embryogenesis.
    Mech Dev. 1993 Oct;43(2-3):87-100 PMID: 8297791
  42. Sequence of C. elegans lag-2 reveals a cell-signalling domain shared with Delta and Serrate of Drosophila.
    Nature. 1994 Mar 10;368(6467):150-4 PMID: 8139658
  43. A member of the Notch group of interacting loci, deltex encodes a cytoplasmic basic protein.
    Genetics. 1994 Feb;136(2):585-96 PMID: 8150285
  44. Recognition sequence of a highly conserved DNA binding protein RBP-J kappa.
    Nucleic Acids Res. 1994 Mar 25;22(6):965-71 PMID: 8152928
  45. The maternal genes apx-1 and glp-1 and establishment of dorsal-ventral polarity in the early C. elegans embryo.
    Cell. 1994 Apr 8;77(1):95-106 PMID: 8156602
  46. Cytosolic interaction between deltex and Notch ankyrin repeats implicates deltex in the Notch signaling pathway.
    Development. 1994 Mar;120(3):473-81 PMID: 8162848
  47. The recombination signal sequence-binding protein RBP-2N functions as a transcriptional repressor.
    Mol Cell Biol. 1994 May;14(5):3310-9 PMID: 8164682
  48. Mediation of Epstein-Barr virus EBNA2 transactivation by recombination signal-binding protein J kappa.
    Science. 1994 Jul 1;265(5168):92-5 PMID: 8016657
  49. The Epstein-Barr virus nuclear antigen 2 transactivator is directed to response elements by the J kappa recombination signal binding protein.
    Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7568-72 PMID: 8052621
  50. EBNA-2 upregulation of Epstein-Barr virus latency promoters and the cellular CD23 promoter utilizes a common targeting intermediate, CBF1.
    J Virol. 1994 Sep;68(9):5375-83 PMID: 8057421
  51. Assembly of recombinant TFIID reveals differential coactivator requirements for distinct transcriptional activators.
    Cell. 1994 Oct 7;79(1):93-105 PMID: 7923382
  52. Notch is required for wingless signaling in the epidermis of Drosophila.
    Cell. 1994 Oct 21;79(2):259-72 PMID: 7954794
  53. The intracellular domain of mouse Notch: a constitutively activated repressor of myogenesis directed at the basic helix-loop-helix region of MyoD.
    Development. 1994 Sep;120(9):2385-96 PMID: 7956819
  54. An activated Notch suppresses neurogenesis and myogenesis but not gliogenesis in mammalian cells.
    Development. 1994 Sep;120(9):2421-30 PMID: 7956822
  55. Epstein-Barr virus nuclear antigen 2 exerts its transactivating function through interaction with recombination signal binding protein RBP-J kappa, the homologue of Drosophila Suppressor of Hairless.
    EMBO J. 1994 Oct 17;13(20):4973-82 PMID: 7957063
  56. Inhibition of the DNA-binding activity of Drosophila suppressor of hairless and of its human homolog, KBF2/RBP-J kappa, by direct protein-protein interaction with Drosophila hairless.
    Genes Dev. 1994 Oct 15;8(20):2491-503 PMID: 7958912
  57. The human J kappa recombination signal sequence binding protein (RBP-J kappa) targets the Epstein-Barr virus EBNA2 protein to its DNA responsive elements.
    EMBO J. 1994 Dec 1;13(23):5633-8 PMID: 7988560
  58. Reciprocal changes in expression of the receptor lin-12 and its ligand lag-2 prior to commitment in a C. elegans cell fate decision.
    Cell. 1994 Dec 30;79(7):1187-98 PMID: 8001154
  59. Jagged: a mammalian ligand that activates Notch1.
    Cell. 1995 Mar 24;80(6):909-17 PMID: 7697721
  60. Notch signaling.
    Science. 1995 Apr 14;268(5208):225-32 PMID: 7716513
  61. Masking of the CBF1/RBPJ kappa transcriptional repression domain by Epstein-Barr virus EBNA2.
    Science. 1995 Apr 28;268(5210):560-3 PMID: 7725102
  62. Primary neurogenesis in Xenopus embryos regulated by a homologue of the Drosophila neurogenic gene Delta.
    Nature. 1995 Jun 29;375(6534):761-6 PMID: 7596407
  63. Alterations in Notch signaling in neoplastic lesions of the human cervix.
    Proc Natl Acad Sci U S A. 1995 Jul 3;92(14):6414-8 PMID: 7604005
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.16.3.952
Published
1996-03-00
Pages
952-9
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC231077
Subset
IM
Grants
NCI NIH HHS · R01 CA42245 · United States
External Links
PubMed source DOI: 10.1128/MCB.16.3.952 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