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EMBO J. Jan 15, 1997; 16(2): 332–342.
PMCID: PMC1169639

Cdk2-dependent phosphorylation of Id2 modulates activity of E2A-related transcription factors.

Abstract

The helix-loop-helix (HLH) protein Id2 is thought to affect the balance between cell growth and differentiation by negatively regulating the function of basic-helix-loop-helix (bHLH) transcription factors. Id2 acts by forming heterodimers that are unable to bind to specific (E-box) DNA sequences. Here we show that this activity can be overcome by phosphorylation of a serine residue within a consensus target site for cyclin-dependent kinases (Cdks). In vitro, Id2 can be phosphorylated by either cyclin E-Cdk2 or cyclin A-Cdk2 but not by cyclin D-dependent kinases. Analogous phosphorylation occurs in serum-stimulated human diploid fibroblasts at a time in late G1 consistent with the appearance of active cyclin E-Cdk2. The phosphorylation of Id2 in these cells correlates with the restoration of a distinct E-box-dependent DNA-binding complex, suggesting that the levels of this complex are modulated by both the abundance and phosphorylation status of Id2. These data provide a link between cyclin-dependent kinases and bHLH transcription factors that may be critical for the regulation of cell proliferation and differentiation.

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Selected References

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  • Ephrussi A, Church GM, Tonegawa S, Gilbert W. B lineage--specific interactions of an immunoglobulin enhancer with cellular factors in vivo. Science. 1985 Jan 11;227(4683):134–140. [PubMed]
  • Ewen ME, Sluss HK, Sherr CJ, Matsushime H, Kato J, Livingston DM. Functional interactions of the retinoblastoma protein with mammalian D-type cyclins. Cell. 1993 May 7;73(3):487–497. [PubMed]
  • Garrell J, Modolell J. The Drosophila extramacrochaetae locus, an antagonist of proneural genes that, like these genes, encodes a helix-loop-helix protein. Cell. 1990 Apr 6;61(1):39–48. [PubMed]
  • Gu W, Schneider JW, Condorelli G, Kaushal S, Mahdavi V, Nadal-Ginard B. Interaction of myogenic factors and the retinoblastoma protein mediates muscle cell commitment and differentiation. Cell. 1993 Feb 12;72(3):309–324. [PubMed]
  • Hara E, Kato T, Nakada S, Sekiya S, Oda K. Subtractive cDNA cloning using oligo(dT)30-latex and PCR: isolation of cDNA clones specific to undifferentiated human embryonal carcinoma cells. Nucleic Acids Res. 1991 Dec;19(25):7097–7104. [PMC free article] [PubMed]
  • Bates S, Bonetta L, MacAllan D, Parry D, Holder A, Dickson C, Peters G. CDK6 (PLSTIRE) and CDK4 (PSK-J3) are a distinct subset of the cyclin-dependent kinases that associate with cyclin D1. Oncogene. 1994 Jan;9(1):71–79. [PubMed]
  • Hara E, Yamaguchi T, Nojima H, Ide T, Campisi J, Okayama H, Oda K. Id-related genes encoding helix-loop-helix proteins are required for G1 progression and are repressed in senescent human fibroblasts. J Biol Chem. 1994 Jan 21;269(3):2139–2145. [PubMed]
  • Begley CG, Aplan PD, Denning SM, Haynes BF, Waldmann TA, Kirsch IR. The gene SCL is expressed during early hematopoiesis and encodes a differentiation-related DNA-binding motif. Proc Natl Acad Sci U S A. 1989 Dec;86(24):10128–10132. [PMC free article] [PubMed]
  • Hatakeyama M, Brill JA, Fink GR, Weinberg RA. Collaboration of G1 cyclins in the functional inactivation of the retinoblastoma protein. Genes Dev. 1994 Aug 1;8(15):1759–1771. [PubMed]
  • Boyle WJ, van der Geer P, Hunter T. Phosphopeptide mapping and phosphoamino acid analysis by two-dimensional separation on thin-layer cellulose plates. Methods Enzymol. 1991;201:110–149. [PubMed]
  • Jan YN, Jan LY. HLH proteins, fly neurogenesis, and vertebrate myogenesis. Cell. 1993 Dec 3;75(5):827–830. [PubMed]
  • Jen Y, Weintraub H, Benezra R. Overexpression of Id protein inhibits the muscle differentiation program: in vivo association of Id with E2A proteins. Genes Dev. 1992 Aug;6(8):1466–1479. [PubMed]
  • Dulić V, Lees E, Reed SI. Association of human cyclin E with a periodic G1-S phase protein kinase. Science. 1992 Sep 25;257(5078):1958–1961. [PubMed]
  • Lam EW, La Thangue NB. DP and E2F proteins: coordinating transcription with cell cycle progression. Curr Opin Cell Biol. 1994 Dec;6(6):859–866. [PubMed]
  • Lasorella A, Iavarone A, Israel MA. Id2 specifically alters regulation of the cell cycle by tumor suppressor proteins. Mol Cell Biol. 1996 Jun;16(6):2570–2578. [PMC free article] [PubMed]
  • Ellenberger T, Fass D, Arnaud M, Harrison SC. Crystal structure of transcription factor E47: E-box recognition by a basic region helix-loop-helix dimer. Genes Dev. 1994 Apr 15;8(8):970–980. [PubMed]
  • Lassar AB, Buskin JN, Lockshon D, Davis RL, Apone S, Hauschka SD, Weintraub H. MyoD is a sequence-specific DNA binding protein requiring a region of myc homology to bind to the muscle creatine kinase enhancer. Cell. 1989 Sep 8;58(5):823–831. [PubMed]
  • Ellis HM, Spann DR, Posakony JW. extramacrochaetae, a negative regulator of sensory organ development in Drosophila, defines a new class of helix-loop-helix proteins. Cell. 1990 Apr 6;61(1):27–38. [PubMed]
  • Lassar AB, Davis RL, Wright WE, Kadesch T, Murre C, Voronova A, Baltimore D, Weintraub H. Functional activity of myogenic HLH proteins requires hetero-oligomerization with E12/E47-like proteins in vivo. Cell. 1991 Jul 26;66(2):305–315. [PubMed]
  • Sorrentino V, Pepperkok R, Davis RL, Ansorge W, Philipson L. Cell proliferation inhibited by MyoD1 independently of myogenic differentiation. Nature. 1990 Jun 28;345(6278):813–815. [PubMed]
  • Le Jossic C, Ilyin GP, Loyer P, Glaise D, Cariou S, Guguen-Guillouzo C. Expression of helix-loop-helix factor Id-1 is dependent on the hepatocyte proliferation and differentiation status in rat liver and in primary culture. Cancer Res. 1994 Dec 1;54(23):6065–6068. [PubMed]
  • Staudinger J, Perry M, Elledge SJ, Olson EN. Interactions among vertebrate helix-loop-helix proteins in yeast using the two-hybrid system. J Biol Chem. 1993 Mar 5;268(7):4608–4611. [PubMed]
  • Sun XH. Constitutive expression of the Id1 gene impairs mouse B cell development. Cell. 1994 Dec 2;79(5):893–900. [PubMed]
  • Sun XH, Copeland NG, Jenkins NA, Baltimore D. Id proteins Id1 and Id2 selectively inhibit DNA binding by one class of helix-loop-helix proteins. Mol Cell Biol. 1991 Nov;11(11):5603–5611. [PMC free article] [PubMed]
  • Weinberg RA. The retinoblastoma protein and cell cycle control. Cell. 1995 May 5;81(3):323–330. [PubMed]
  • Mellentin JD, Smith SD, Cleary ML. lyl-1, a novel gene altered by chromosomal translocation in T cell leukemia, codes for a protein with a helix-loop-helix DNA binding motif. Cell. 1989 Jul 14;58(1):77–83. [PubMed]
  • Weintraub H. The MyoD family and myogenesis: redundancy, networks, and thresholds. Cell. 1993 Dec 31;75(7):1241–1244. [PubMed]
  • Meyerson M, Harlow E. Identification of G1 kinase activity for cdk6, a novel cyclin D partner. Mol Cell Biol. 1994 Mar;14(3):2077–2086. [PMC free article] [PubMed]
  • Wilson R, Mohun T. XIdx, a dominant negative regulator of bHLH function in early Xenopus embryos. Mech Dev. 1995 Feb;49(3):211–222. [PubMed]
  • Zhang H, Reynaud S, Kloc M, Etkin LD, Spohr G. Id gene activity during Xenopus embryogenesis. Mech Dev. 1995 Apr;50(2-3):119–130. [PubMed]
  • Murre C, McCaw PS, Vaessin H, Caudy M, Jan LY, Jan YN, Cabrera CV, Buskin JN, Hauschka SD, Lassar AB, et al. Interactions between heterologous helix-loop-helix proteins generate complexes that bind specifically to a common DNA sequence. Cell. 1989 Aug 11;58(3):537–544. [PubMed]
  • Zhang Y, Babin J, Feldhaus AL, Singh H, Sharp PA, Bina M. HTF4: a new human helix-loop-helix protein. Nucleic Acids Res. 1991 Aug 25;19(16):4555–4555. [PMC free article] [PubMed]
  • Nagata Y, Shoji W, Obinata M, Todokoro K. Phosphorylation of helix-loop-helix proteins ID1, ID2 and ID3. Biochem Biophys Res Commun. 1995 Feb 27;207(3):916–926. [PubMed]
  • Naya FJ, Stellrecht CM, Tsai MJ. Tissue-specific regulation of the insulin gene by a novel basic helix-loop-helix transcription factor. Genes Dev. 1995 Apr 15;9(8):1009–1019. [PubMed]
  • Ohashi M, Aizawa S, Ooka H, Ohsawa T, Kaji K, Kondo H, Kobayashi T, Noumura T, Matsuo M, Mitsui Y, et al. A new human diploid cell strain, TIG-1, for the research on cellular aging. Exp Gerontol. 1980;15(2):121–133. [PubMed]
  • Ohtsubo M, Theodoras AM, Schumacher J, Roberts JM, Pagano M. Human cyclin E, a nuclear protein essential for the G1-to-S phase transition. Mol Cell Biol. 1995 May;15(5):2612–2624. [PMC free article] [PubMed]
  • Olson EN. MyoD family: a paradigm for development? Genes Dev. 1990 Sep;4(9):1454–1461. [PubMed]
  • Pagliuca A, Bartoli PC, Saccone S, Della Valle G, Lania L. Molecular cloning of ID4, a novel dominant negative helix-loop-helix human gene on chromosome 6p21.3-p22. Genomics. 1995 May 1;27(1):200–203. [PubMed]
  • Parry D, Bates S, Mann DJ, Peters G. Lack of cyclin D-Cdk complexes in Rb-negative cells correlates with high levels of p16INK4/MTS1 tumour suppressor gene product. EMBO J. 1995 Feb 1;14(3):503–511. [PMC free article] [PubMed]
  • Peters G. The D-type cyclins and their role in tumorigenesis. J Cell Sci Suppl. 1994;18:89–96. [PubMed]
  • Peverali FA, Ramqvist T, Saffrich R, Pepperkok R, Barone MV, Philipson L. Regulation of G1 progression by E2A and Id helix-loop-helix proteins. EMBO J. 1994 Sep 15;13(18):4291–4301. [PMC free article] [PubMed]
  • Resnitzky D, Reed SI. Different roles for cyclins D1 and E in regulation of the G1-to-S transition. Mol Cell Biol. 1995 Jul;15(7):3463–3469. [PMC free article] [PubMed]
  • Riechmann V, van Crüchten I, Sablitzky F. The expression pattern of Id4, a novel dominant negative helix-loop-helix protein, is distinct from Id1, Id2 and Id3. Nucleic Acids Res. 1994 Mar 11;22(5):749–755. [PMC free article] [PubMed]
  • Shen CP, Kadesch T. B-cell-specific DNA binding by an E47 homodimer. Mol Cell Biol. 1995 Aug;15(8):4518–4524. [PMC free article] [PubMed]
  • Sherr CJ. G1 phase progression: cycling on cue. Cell. 1994 Nov 18;79(4):551–555. [PubMed]
  • Shoji W, Yamamoto T, Obinata M. The helix-loop-helix protein Id inhibits differentiation of murine erythroleukemia cells. J Biol Chem. 1994 Feb 18;269(7):5078–5084. [PubMed]
  • Shoji W, Inoue T, Yamamoto T, Obinata M. MIDA1, a protein associated with Id, regulates cell growth. J Biol Chem. 1995 Oct 20;270(42):24818–24825. [PubMed]
  • Skerjanc IS, Truong J, Filion P, McBurney MW. A splice variant of the ITF-2 transcript encodes a transcription factor that inhibits MyoD activity. J Biol Chem. 1996 Feb 16;271(7):3555–3561. [PubMed]
  • Smith DB, Johnson KS. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. [PubMed]

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