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PMID: 9199321 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Specific regulation of E2F family members by cyclin-dependent kinases.

Molecular and cellular biology ·Vol. 17 ·No. 7 ·1997-07-00 ·Pages 3867-75

Dynlacht BD, Moberg K, Lees JA, Harlow E, Zhu L

Abstract

The transcription factor E2F-1 interacts stably with cyclin A via a small domain near its amino terminus and is negatively regulated by the cyclin A-dependent kinases. Thus, the activities of E2F, a family of transcription factors involved in cell proliferation, are regulated by at least two types of cell growth regulators: the retinoblastoma protein family and the cyclin-dependent kinase family. To investigate further the regulation of E2F by cyclin-dependent kinases, we have extended our studies to include additional cyclins and E2F family members. Using purified components in an in vitro system, we show that the E2F-1-DP-1 heterodimer, the functionally active form of the E2F activity, is not a substrate for the active cyclin D-dependent kinases but is efficiently phosphorylated by the cyclin B-dependent kinases, which do not form stable complexes with the E2F-1-DP-1 heterodimer. Phosphorylation of the E2F-1-DP-1 heterodimer by cyclin B-dependent kinases, however, did not result in down-regulation of its DNA-binding activity, as is readily seen after phosphorylation by cyclin A-dependent kinases, suggesting that phosphorylation per se is not sufficient to regulate E2F DNA-binding activity. Furthermore, heterodimers containing E2F-4, a family member lacking the cyclin A binding domain found in E2F-1, are not efficiently phosphorylated or functionally down-regulated by cyclin A-dependent kinases. However, addition of the E2F-1 cyclin A binding domain to E2F-4 conferred cyclin A-dependent kinase-mediated down-regulation of the E2F-4-DP-1 heterodimer. Thus, both enzymatic phosphorylation and stable physical interaction are necessary for the specific regulation of E2F family members by cyclin-dependent kinases.

MeSH Terms
CDC2-CDC28 Kinases Carrier Proteins Cell Cycle Cell Cycle Proteins Cell-Free System Cyclin-Dependent Kinase 2 Cyclin-Dependent Kinases/metabolism Cyclins/metabolism DNA-Binding Proteins/metabolism E2F Transcription Factors E2F1 Transcription Factor Peptide Mapping Phosphopeptides/analysis Protein Binding Protein Serine-Threonine Kinases/metabolism Recombinant Proteins Retinoblastoma-Binding Protein 1 Structure-Activity Relationship Substrate Specificity Transcription Factors/physiology
Chemicals
Carrier Proteins Cell Cycle Proteins Cyclins DNA-Binding Proteins E2F Transcription Factors E2F1 Transcription Factor Phosphopeptides Recombinant Proteins Retinoblastoma-Binding Protein 1 Transcription Factors Protein Serine-Threonine Kinases CDC2-CDC28 Kinases Cyclin-Dependent Kinase 2 Cyclin-Dependent Kinases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Dynlacht B D
Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts 02138, USA. dynlacht@biosun.harvard.edu
Moberg K
Lees J A
Harlow E
Zhu L
References (53)
53 references, click to expand
  1. A cDNA encoding a pRB-binding protein with properties of the transcription factor E2F.
    Cell. 1992 Jul 24;70(2):337-50 PMID: 1638634
  2. Molecular cloning of cellular genes encoding retinoblastoma-associated proteins: identification of a gene with properties of the transcription factor E2F.
    Mol Cell Biol. 1992 Dec;12(12):5620-31 PMID: 1448092
  3. E2F-1 functions in mice to promote apoptosis and suppress proliferation.
    Cell. 1996 May 17;85(4):549-61 PMID: 8653790
  4. Tumor induction and tissue atrophy in mice lacking E2F-1.
    Cell. 1996 May 17;85(4):537-48 PMID: 8653789
  5. Cloning and characterization of E2F-2, a novel protein with the biochemical properties of transcription factor E2F.
    Mol Cell Biol. 1993 Dec;13(12):7802-12 PMID: 8246995
  6. Identification and properties of an atypical catalytic subunit (p34PSK-J3/cdk4) for mammalian D type G1 cyclins.
    Cell. 1992 Oct 16;71(2):323-34 PMID: 1423597
  7. p107 uses a p21CIP1-related domain to bind cyclin/cdk2 and regulate interactions with E2F.
    Genes Dev. 1995 Jul 15;9(14):1740-52 PMID: 7622038
  8. E2F: a link between the Rb tumor suppressor protein and viral oncoproteins.
    Science. 1992 Oct 16;258(5081):424-9 PMID: 1411535
  9. Cell-cycle inhibition by independent CDK and PCNA binding domains in p21Cip1.
    Nature. 1995 May 11;375(6527):159-61 PMID: 7753174
  10. The transcription factor E2F interacts with the retinoblastoma product and a p107-cyclin A complex in a cell cycle-regulated manner.
    Cell. 1992 Jan 10;68(1):157-66 PMID: 1531040
  11. Cyclin E/cdk2 and cyclin A/cdk2 kinases associate with p107 and E2F in a temporally distinct manner.
    Genes Dev. 1992 Oct;6(10):1874-85 PMID: 1398067
  12. 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
  13. Direct binding of cyclin D to the retinoblastoma gene product (pRb) and pRb phosphorylation by the cyclin D-dependent kinase CDK4.
    Genes Dev. 1993 Mar;7(3):331-42 PMID: 8449399
  14. Differential regulation of E2F transactivation by cyclin/cdk2 complexes.
    Genes Dev. 1994 Aug 1;8(15):1772-86 PMID: 7958856
  15. Multiple members of the E2F transcription factor family are the products of oncogenes.
    Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1357-61 PMID: 7877982
  16. D-type cyclin-dependent kinase activity in mammalian cells.
    Mol Cell Biol. 1994 Mar;14(3):2066-76 PMID: 8114738
  17. Physical interaction of the retinoblastoma protein with human D cyclins.
    Cell. 1993 May 7;73(3):499-511 PMID: 8490963
  18. Crystal structure of the p27Kip1 cyclin-dependent-kinase inhibitor bound to the cyclin A-Cdk2 complex.
    Nature. 1996 Jul 25;382(6589):325-31 PMID: 8684460
  19. The retinoblastoma protein as a transcriptional repressor.
    Trends Cell Biol. 1993 Feb;3(2):43-6 PMID: 14731718
  20. Characterization of p21Cip1/Waf1 peptide domains required for cyclin E/Cdk2 and PCNA interaction.
    Oncogene. 1996 Feb 1;12(3):595-607 PMID: 8637717
  21. Cyclins and cancer. II: Cyclin D and CDK inhibitors come of age.
    Cell. 1994 Nov 18;79(4):573-82 PMID: 7954824
  22. E2F-5, a new E2F family member that interacts with p130 in vivo.
    Mol Cell Biol. 1995 Jun;15(6):3082-9 PMID: 7760804
  23. A- and B-type cyclins differentially modulate substrate specificity of cyclin-cdk complexes.
    EMBO J. 1993 May;12(5):1947-54 PMID: 8491188
  24. Cyclin A-kinase regulation of E2F-1 DNA binding function underlies suppression of an S phase checkpoint.
    Cell. 1995 Dec 29;83(7):1149-58 PMID: 8548802
  25. p53 and E2F-1 cooperate to mediate apoptosis.
    Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3602-6 PMID: 8170954
  26. E2F-4 and E2F-5, two members of the E2F family, are expressed in the early phases of the cell cycle.
    Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):2403-7 PMID: 7892279
  27. Expression cloning of a cDNA encoding a retinoblastoma-binding protein with E2F-like properties.
    Cell. 1992 Jul 24;70(2):351-64 PMID: 1638635
  28. Cyclin-binding motifs are essential for the function of p21CIP1.
    Mol Cell Biol. 1996 Sep;16(9):4673-82 PMID: 8756624
  29. 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
  30. Inhibition of E2F-1 transactivation by direct binding of the retinoblastoma protein.
    Mol Cell Biol. 1993 Oct;13(10):6501-8 PMID: 8413249
  31. Interaction between human cyclin A and adenovirus E1A-associated p107 protein.
    Science. 1992 Jan 3;255(5040):87-90 PMID: 1532458
  32. Deregulated expression of E2F-1 induces S-phase entry and leads to apoptosis.
    Mol Cell Biol. 1994 Dec;14(12):8166-73 PMID: 7969153
  33. Functional interaction between E2F-4 and p130: evidence for distinct mechanisms underlying growth suppression by different retinoblastoma protein family members.
    Genes Dev. 1995 Apr 1;9(7):869-81 PMID: 7705662
  34. Transcriptional regulation during the mammalian cell cycle.
    Trends Genet. 1995 May;11(5):173-8 PMID: 7785076
  35. Overexpression of E2F-1 in rat embryo fibroblasts leads to neoplastic transformation.
    EMBO J. 1994 Jul 15;13(14):3329-38 PMID: 8045262
  36. Deregulated transcription factor E2F-1 expression leads to S-phase entry and p53-mediated apoptosis.
    Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):10918-22 PMID: 7971984
  37. A cyclin A-protein kinase complex possesses sequence-specific DNA binding activity: p33cdk2 is a component of the E2F-cyclin A complex.
    Cell. 1992 Jan 10;68(1):167-76 PMID: 1310073
  38. Functional interactions of the retinoblastoma protein with mammalian D-type cyclins.
    Cell. 1993 May 7;73(3):487-97 PMID: 8343202
  39. Independent binding of the retinoblastoma protein and p107 to the transcription factor E2F.
    Nature. 1992 Jan 9;355(6356):176-9 PMID: 1530885
  40. Distinct roles for cyclin-dependent kinases in cell cycle control.
    Science. 1993 Dec 24;262(5142):2050-4 PMID: 8266103
  41. E2F-4, a new member of the E2F transcription factor family, interacts with p107.
    Genes Dev. 1994 Nov 15;8(22):2665-79 PMID: 7958924
  42. Cyclin A/CDK2 binds directly to E2F-1 and inhibits the DNA-binding activity of E2F-1/DP-1 by phosphorylation.
    Mol Cell Biol. 1994 Dec;14(12):8420-31 PMID: 7969176
  43. Negative regulation of the growth-promoting transcription factor E2F-1 by a stably bound cyclin A-dependent protein kinase.
    Cell. 1994 Jul 15;78(1):161-72 PMID: 8033208
  44. Expression of transcription factor E2F1 induces quiescent cells to enter S phase.
    Nature. 1993 Sep 23;365(6444):349-52 PMID: 8377827
  45. Transcription factor E2F binds DNA as a heterodimer.
    Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3525-9 PMID: 8475102
  46. pRB, p107 and the regulation of the E2F transcription factor.
    J Cell Sci Suppl. 1994;18:81-7 PMID: 7883798
  47. Activation of human cyclin-dependent kinases in vitro.
    Mol Biol Cell. 1992 May;3(5):571-82 PMID: 1535244
  48. Analysis of wild-type and mutant p21WAF-1 gene activities.
    Mol Cell Biol. 1996 Apr;16(4):1786-93 PMID: 8657154
  49. E2F1 overexpression in quiescent fibroblasts leads to induction of cellular DNA synthesis and apoptosis.
    J Virol. 1995 Apr;69(4):2491-500 PMID: 7884898
  50. 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
  51. Identification of G1 kinase activity for cdk6, a novel cyclin D partner.
    Mol Cell Biol. 1994 Mar;14(3):2077-86 PMID: 8114739
  52. E2F-4, a new member of the E2F gene family, has oncogenic activity and associates with p107 in vivo.
    Genes Dev. 1994 Nov 15;8(22):2680-90 PMID: 7958925
  53. Retinoblastoma protein switches the E2F site from positive to negative element.
    Nature. 1992 Jul 16;358(6383):259-61 PMID: 1321348
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.17.7.3867
Email
dynlacht@biosun.harvard.edu
Published
1997-07-00
Pages
3867-75
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC232239
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.17.7.3867 PMC Full Text View journal details
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