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PMID: 12502741 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Structural basis for the recognition of the E2F transactivation domain by the retinoblastoma tumor suppressor.

Genes & development ·Vol. 16 ·No. 24 ·2002-12-15 ·Pages 3199-212

Lee C, Chang JH, Lee HS, Cho Y

Abstract

Repression of E2F transcription activity by the retinoblastoma (Rb) tumor suppressor through its interaction with the transactivation domain of the E2F transcription factor is one of the central features of G1/S arrest in the mammalian cell cycle. Deregulation of the Rb-E2F interaction results in hyperproliferation, lack of differentiation, and apoptosis, and can lead to cancer. The 2.2-A crystal structure of the Rb pocket complexed with an 18-residue transactivation-domain peptide of E2F-2 reveals that the boomerang-shaped peptide binds to the highly conserved interface between the A-box and the B-box of the Rb pocket in a bipartite manner. The N-terminal segment of the E2F-2 peptide in an extended beta-strand-like structure interacts with helices from the conserved groove at the A-B interface, whereas the C-terminal segment, which contains one 3(10) helix, binds to a groove mainly formed by A-box helices. The flexibility in the middle of the E2F-2 peptide is essential for the tight association of E2F to the Rb pocket. The binding of Rb to the E2F-2 peptide conceals several conserved residues that are crucial for transcription activation of E2F. We provide the structural basis for the Rb-mediated repression of E2F transcription activity without the requirement of histone-modifying enzymes.

MeSH Terms
Cell Cycle Proteins Conserved Sequence Crystallography, X-Ray DNA-Binding Proteins E2F Transcription Factors E2F2 Transcription Factor Humans Molecular Sequence Data Mutation Peptide Fragments/genetics,metabolism Protein Binding Protein Structure, Tertiary Retinoblastoma Protein/genetics,metabolism Sequence Homology, Amino Acid Surface Plasmon Resonance Transcription Factors/genetics,metabolism Transcriptional Activation
Chemicals
Cell Cycle Proteins DNA-Binding Proteins E2F Transcription Factors E2F2 Transcription Factor E2F2 protein, human Peptide Fragments Retinoblastoma Protein Transcription Factors
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Lee Changwook
National Creative Research Center for Structural Biology and Department of Life Science, Pohang University of Science and Technology, San 31, KyungBook, South Korea.
Chang Jeong Ho
Lee Hyun Sook
Cho Yunje
References (74)
74 references, click to expand
  1. Mechanism of transcriptional repression of E2F by the retinoblastoma tumor suppressor protein.
    Mol Cell. 1999 Feb;3(2):195-205 PMID: 10078202
  2. Active transcriptional repression by the Rb-E2F complex mediates G1 arrest triggered by p16INK4a, TGFbeta, and contact inhibition.
    Cell. 1999 Apr 2;97(1):53-61 PMID: 10199402
  3. Activation of the murine dihydrofolate reductase promoter by E2F1. A requirement for CBP recruitment.
    J Biol Chem. 1999 May 28;274(22):15883-91 PMID: 10336493
  4. Cdk phosphorylation triggers sequential intramolecular interactions that progressively block Rb functions as cells move through G1.
    Cell. 1999 Sep 17;98(6):859-69 PMID: 10499802
  5. Exit from G1 and S phase of the cell cycle is regulated by repressor complexes containing HDAC-Rb-hSWI/SNF and Rb-hSWI/SNF.
    Cell. 2000 Mar 31;101(1):79-89 PMID: 10778858
  6. Rb function in cell-cycle regulation and apoptosis.
    Nat Cell Biol. 2000 Apr;2(4):E65-7 PMID: 10783254
  7. The Rb/E2F pathway: expanding roles and emerging paradigms.
    Genes Dev. 2000 Oct 1;14(19):2393-409 PMID: 11018009
  8. Role for the p53 homologue p73 in E2F-1-induced apoptosis.
    Nature. 2000 Oct 5;407(6804):645-8 PMID: 11034215
  9. Structural basis for the inactivation of retinoblastoma tumor suppressor by SV40 large T antigen.
    EMBO J. 2001 Jan 15;20(1-2):295-304 PMID: 11226179
  10. The Rb/E2F pathway and cancer.
    Hum Mol Genet. 2001 Apr;10(7):699-703 PMID: 11257102
  11. Rb targets histone H3 methylation and HP1 to promoters.
    Nature. 2001 Aug 2;412(6846):561-5 PMID: 11484059
  12. Inactivation of the retinoblastoma tumor suppressor induces apoptosis protease-activating factor-1 dependent and independent apoptotic pathways during embryogenesis.
    Cancer Res. 2001 Dec 1;61(23):8395-400 PMID: 11731416
  13. Interactions of SV40 large T antigen and other viral proteins with retinoblastoma tumour suppressor.
    Rev Med Virol. 2002 Mar-Apr;12(2):81-92 PMID: 11921304
  14. Structural basis for recruitment of CBP/p300 by hypoxia-inducible factor-1 alpha.
    Proc Natl Acad Sci U S A. 2002 Apr 16;99(8):5367-72 PMID: 11959990
  15. Adenovirus E1A, simian virus 40 tumor antigen, and human papillomavirus E7 protein share the capacity to disrupt the interaction between transcription factor E2F and the retinoblastoma gene product.
    Proc Natl Acad Sci U S A. 1992 May 15;89(10):4549-53 PMID: 1316611
  16. Oncogenic point mutations in exon 20 of the RB1 gene in families showing incomplete penetrance and mild expression of the retinoblastoma phenotype.
    Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6177-81 PMID: 1352883
  17. Target enzyme recognition by calmodulin: 2.4 A structure of a calmodulin-peptide complex.
    Science. 1992 Aug 28;257(5074):1251-5 PMID: 1519061
  18. The CCP4 suite: programs for protein crystallography.
    Acta Crystallogr D Biol Crystallogr. 1994 Sep 1;50(Pt 5):760-3 PMID: 15299374
  19. The interaction of RB with E2F coincides with an inhibition of the transcriptional activity of E2F.
    Genes Dev. 1992 Feb;6(2):177-85 PMID: 1531329
  20. Thymidine kinase transcription is regulated at G1/S phase by a complex that contains retinoblastoma-like protein and a cdc2 kinase.
    Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3256-60 PMID: 1565617
  21. Cell cycle regulation of the human cdc2 gene.
    EMBO J. 1992 May;11(5):1797-804 PMID: 1582409
  22. A cDNA encoding a pRB-binding protein with properties of the transcription factor E2F.
    Cell. 1992 Jul 24;70(2):337-50 PMID: 1638634
  23. Expression cloning of a cDNA encoding a retinoblastoma-binding protein with E2F-like properties.
    Cell. 1992 Jul 24;70(2):351-64 PMID: 1638635
  24. Nonfunctional mutants of the retinoblastoma protein are characterized by defects in phosphorylation, viral oncoprotein association, and nuclear tethering.
    Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3033-7 PMID: 1826560
  25. The retinoblastoma protein copurifies with E2F-I, an E1A-regulated inhibitor of the transcription factor E2F.
    Cell. 1991 Jun 14;65(6):1063-72 PMID: 1828393
  26. Human DNA polymerase alpha gene: sequences controlling expression in cycling and serum-stimulated cells.
    Mol Cell Biol. 1991 Apr;11(4):2081-95 PMID: 2005899
  27. Improved methods for building protein models in electron density maps and the location of errors in these models.
    Acta Crystallogr A. 1991 Mar 1;47 ( Pt 2):110-9 PMID: 2025413
  28. The regions of the retinoblastoma protein needed for binding to adenovirus E1A or SV40 large T antigen are common sites for mutations.
    EMBO J. 1990 Apr;9(4):1147-55 PMID: 2138977
  29. A role for the adenovirus inducible E2F transcription factor in a proliferation dependent signal transduction pathway.
    EMBO J. 1990 Jul;9(7):2179-84 PMID: 2141565
  30. The retinoblastoma susceptibility gene product undergoes cell cycle-dependent dephosphorylation and binding to and release from SV40 large T.
    Cell. 1990 Feb 9;60(3):387-96 PMID: 2154332
  31. Definition of the minimal simian virus 40 large T antigen- and adenovirus E1A-binding domain in the retinoblastoma gene product.
    Mol Cell Biol. 1990 Jul;10(7):3761-9 PMID: 2162480
  32. Two distinct and frequently mutated regions of retinoblastoma protein are required for binding to SV40 T antigen.
    EMBO J. 1990 Jun;9(6):1815-22 PMID: 2189724
  33. E1A-dependent trans-activation of the human MYC promoter is mediated by the E2F factor.
    Proc Natl Acad Sci U S A. 1989 May;86(10):3594-8 PMID: 2524830
  34. Transcription factor E2F is required for efficient expression of the hamster dihydrofolate reductase gene in vitro and in vivo.
    Mol Cell Biol. 1989 Nov;9(11):4994-5002 PMID: 2601705
  35. The product of the retinoblastoma susceptibility gene has properties of a cell cycle regulatory element.
    Cell. 1989 Sep 22;58(6):1085-95 PMID: 2673542
  36. Nuclear factor E2F mediates basic transcription and trans-activation by E1a of the human MYC promoter.
    Genes Dev. 1989 Apr;3(4):527-36 PMID: 2721961
  37. Processing of X-ray diffraction data collected in oscillation mode.
    Methods Enzymol. 1997;276:307-26 PMID: 27754618
  38. Structure of the human class I histocompatibility antigen, HLA-A2.
    Nature. 1987 Oct 8-14;329(6139):506-12 PMID: 3309677
  39. Cell cycle regulation of the cyclin A gene promoter is mediated by a variant E2F site.
    Proc Natl Acad Sci U S A. 1995 Nov 21;92(24):11264-8 PMID: 7479977
  40. Mechanism of active transcriptional repression by the retinoblastoma protein.
    Nature. 1995 Jun 29;375(6534):812-5 PMID: 7596417
  41. Cellular targets for activation by the E2F1 transcription factor include DNA synthesis- and G1/S-regulatory genes.
    Mol Cell Biol. 1995 Aug;15(8):4215-24 PMID: 7623816
  42. The retinoblastoma protein and cell cycle control.
    Cell. 1995 May 5;81(3):323-30 PMID: 7736585
  43. E2F-5, a new E2F family member that interacts with p130 in vivo.
    Mol Cell Biol. 1995 Jun;15(6):3082-9 PMID: 7760804
  44. The role of cellular transcription factor E2F in the regulation of cdc2 mRNA expression and cell cycle control of human hematopoietic cells.
    J Biol Chem. 1994 Oct 21;269(42):26249-58 PMID: 7929342
  45. Differential regulation of E2F transactivation by cyclin/cdk2 complexes.
    Genes Dev. 1994 Aug 1;8(15):1772-86 PMID: 7958856
  46. Partial inactivation of the RB product in a family with incomplete penetrance of familial retinoblastoma and benign retinal tumors.
    Oncogene. 1994 May;9(5):1321-6 PMID: 8152792
  47. The retinoblastoma protein binds to a family of E2F transcription factors.
    Mol Cell Biol. 1993 Dec;13(12):7813-25 PMID: 8246996
  48. Binding to DNA and the retinoblastoma gene product promoted by complex formation of different E2F family members.
    Science. 1993 Dec 3;262(5139):1557-60 PMID: 8248803
  49. The retinoblastoma protein binds E2F residues required for activation in vivo and TBP binding in vitro.
    Nucleic Acids Res. 1993 Nov 11;21(22):4998-5004 PMID: 8255752
  50. E2F-1-mediated transactivation is inhibited by complex formation with the retinoblastoma susceptibility gene product.
    Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):6914-8 PMID: 8346196
  51. Heterodimerization of the transcription factors E2F-1 and DP-1 leads to cooperative trans-activation.
    Genes Dev. 1993 Oct;7(10):1850-61 PMID: 8405995
  52. A genetic analysis of the E2F1 gene distinguishes regulation by Rb, p107, and adenovirus E4.
    Mol Cell Biol. 1993 Oct;13(10):6314-25 PMID: 8413230
  53. Inhibition of E2F-1 transactivation by direct binding of the retinoblastoma protein.
    Mol Cell Biol. 1993 Oct;13(10):6501-8 PMID: 8413249
  54. Disruption of RB/E2F-1 interaction by single point mutations in E2F-1 enhances S-phase entry and apoptosis.
    Proc Natl Acad Sci U S A. 1996 Jan 23;93(2):679-84 PMID: 8570615
  55. A unique role for the Rb protein in controlling E2F accumulation during cell growth and differentiation.
    Proc Natl Acad Sci U S A. 1996 Apr 16;93(8):3215-20 PMID: 8622916
  56. E2F1 and E1A(12S) have a homologous activation domain regulated by RB and CBP.
    Proc Natl Acad Sci U S A. 1996 Feb 20;93(4):1439-42 PMID: 8643650
  57. Tumor induction and tissue atrophy in mice lacking E2F-1.
    Cell. 1996 May 17;85(4):537-48 PMID: 8653789
  58. E2F-1 functions in mice to promote apoptosis and suppress proliferation.
    Cell. 1996 May 17;85(4):549-61 PMID: 8653790
  59. E2F-4 switches from p130 to p107 and pRB in response to cell cycle reentry.
    Mol Cell Biol. 1996 Apr;16(4):1436-49 PMID: 8657117
  60. Structure of the MDM2 oncoprotein bound to the p53 tumor suppressor transactivation domain.
    Science. 1996 Nov 8;274(5289):948-53 PMID: 8875929
  61. The CBP co-activator stimulates E2F1/DP1 activity.
    Nucleic Acids Res. 1996 Nov 1;24(21):4139-45 PMID: 8932363
  62. Induced alpha helix in the VP16 activation domain upon binding to a human TAF.
    Science. 1997 Aug 29;277(5330):1310-3 PMID: 9271577
  63. Regulation of transcription by proteins that control the cell cycle.
    Nature. 1997 Sep 11;389(6647):149-52 PMID: 9296491
  64. Hypo-phosphorylation of the retinoblastoma protein (pRb) by cyclin D:Cdk4/6 complexes results in active pRb.
    Proc Natl Acad Sci U S A. 1997 Sep 30;94(20):10699-704 PMID: 9380698
  65. Modular organization of the E2F1 activation domain and its interaction with general transcription factors TBP and TFIIH.
    Oncogene. 1997 Nov 27;15(22):2643-58 PMID: 9400991
  66. Solution structure of the KIX domain of CBP bound to the transactivation domain of CREB: a model for activator:coactivator interactions.
    Cell. 1997 Dec 12;91(6):741-52 PMID: 9413984
  67. Functional inactivation of the retinoblastoma protein requires sequential modification by at least two distinct cyclin-cdk complexes.
    Mol Cell Biol. 1998 Feb;18(2):753-61 PMID: 9447971
  68. Retinoblastoma protein recruits histone deacetylase to repress transcription.
    Nature. 1998 Feb 5;391(6667):597-601 PMID: 9468139
  69. Rb interacts with histone deacetylase to repress transcription.
    Cell. 1998 Feb 20;92(4):463-73 PMID: 9491888
  70. Structure of the retinoblastoma tumour-suppressor pocket domain bound to a peptide from HPV E7.
    Nature. 1998 Feb 26;391(6670):859-65 PMID: 9495340
  71. Loss of E2F-1 reduces tumorigenesis and extends the lifespan of Rb1(+/-)mice.
    Nat Genet. 1998 Apr;18(4):360-4 PMID: 9537419
  72. Increased E2F1 activity induces skin tumors in mice heterozygous and nullizygous for p53.
    Proc Natl Acad Sci U S A. 1998 Jul 21;95(15):8858-63 PMID: 9671769
  73. The regulation of E2F by pRB-family proteins.
    Genes Dev. 1998 Aug 1;12(15):2245-62 PMID: 9694791
  74. Crystallography & NMR system: A new software suite for macromolecular structure determination.
    Acta Crystallogr D Biol Crystallogr. 1998 Sep 1;54(Pt 5):905-21 PMID: 9757107
Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.1046102
Published
2002-12-15
Pages
3199-212
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC187509
Subset
IM
Databases
PDB
1N4M
External Links
PubMed source DOI: 10.1101/gad.1046102 PMC Full Text View journal details
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