• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of molcellbPermissionsJournals.ASM.orgJournalMCB ArticleJournal InfoAuthorsReviewers
Mol Cell Biol. Nov 1997; 17(11): 6563–6573.
PMCID: PMC232510

The basic domain of myogenic basic helix-loop-helix (bHLH) proteins is the novel target for direct inhibition by another bHLH protein, Twist.

Abstract

In vertebrates, the basic helix-loop-helix (bHLH) protein Twist may be involved in the negative regulation of cellular determination and in the differentiation of several lineages, including myogenesis, osteogenesis, and neurogenesis. Although it has been shown that mouse twist (M-Twist) (i) sequesters E proteins, thus preventing formation of myogenic E protein-MyoD complexes and (ii) inhibits the MEF2 transcription factor, a cofactor of myogenic bHLH proteins, overexpression of E proteins and MEF2 failed to rescue the inhibitory effects of M-Twist on MyoD. We report here that M-Twist physically interacts with the myogenic bHLH proteins in vitro and in vivo and that this interaction is required for the inhibition of MyoD by M-Twist. In contrast to the conventional HLH-HLH domain interaction formed in the MyoD/E12 heterodimer, this novel type of interaction uses the basic domains of the two proteins. While the MyoD HLH domain without the basic domain failed to interact with M-Twist, a MyoD peptide containing only the basic and helix 1 regions was sufficient to interact with M-Twist, suggesting that the basic domain contacts M-Twist. The replacement of three arginine residues by alanines in the M-Twist basic domain was sufficient to abolish both the binding and inhibition of MyoD by M-Twist, while the domain retained other M-Twist functions such as heterodimerization with an E protein and inhibition of MEF2 transactivation. These findings demonstrate that M-Twist interacts with MyoD through the basic domains, thereby inhibiting MyoD.

Full Text

The Full Text of this article is available as a PDF (1.3M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Hebrok M, Füchtbauer A, Füchtbauer EM. Repression of muscle-specific gene activation by the murine Twist protein. Exp Cell Res. 1997 May 1;232(2):295–303. [PubMed]
  • Anderson DJ. Stem cells and transcription factors in the development of the mammalian neural crest. FASEB J. 1994 Jul;8(10):707–713. [PubMed]
  • Anthony-Cahill SJ, Benfield PA, Fairman R, Wasserman ZR, Brenner SL, Stafford WF, 3rd, Altenbach C, Hubbell WL, DeGrado WF. Molecular characterization of helix-loop-helix peptides. Science. 1992 Feb 21;255(5047):979–983. [PubMed]
  • Artandi SE, Cooper C, Shrivastava A, Calame K. The basic helix-loop-helix-zipper domain of TFE3 mediates enhancer-promoter interaction. Mol Cell Biol. 1994 Dec;14(12):7704–7716. [PMC free article] [PubMed]
  • Bardwell VJ, Treisman R. The POZ domain: a conserved protein-protein interaction motif. Genes Dev. 1994 Jul 15;8(14):1664–1677. [PubMed]
  • Baylies MK, Bate M. twist: a myogenic switch in Drosophila. Science. 1996 Jun 7;272(5267):1481–1484. [PubMed]
  • Benezra R, Davis RL, Lassar A, Tapscott S, Thayer M, Lockshon D, Weintraub H. Id: a negative regulator of helix-loop-helix DNA binding proteins. Control of terminal myogenic differentiation. Ann N Y Acad Sci. 1990;599:1–11. [PubMed]
  • Bengal E, Flores O, Rangarajan PN, Chen A, Weintraub H, Verma IM. Positive control mutations in the MyoD basic region fail to show cooperative DNA binding and transcriptional activation in vitro. Proc Natl Acad Sci U S A. 1994 Jun 21;91(13):6221–6225. [PMC free article] [PubMed]
  • Bengal E, Ransone L, Scharfmann R, Dwarki VJ, Tapscott SJ, Weintraub H, Verma IM. Functional antagonism between c-Jun and MyoD proteins: a direct physical association. Cell. 1992 Feb 7;68(3):507–519. [PubMed]
  • Bernard O, Cory S, Gerondakis S, Webb E, Adams JM. Sequence of the murine and human cellular myc oncogenes and two modes of myc transcription resulting from chromosome translocation in B lymphoid tumours. EMBO J. 1983;2(12):2375–2383. [PMC free article] [PubMed]
  • Blackwell TK, Weintraub H. Differences and similarities in DNA-binding preferences of MyoD and E2A protein complexes revealed by binding site selection. Science. 1990 Nov 23;250(4984):1104–1110. [PubMed]
  • Blanar MA, Crossley PH, Peters KG, Steingrímsson E, Copeland NG, Jenkins NA, Martin GR, Rutter WJ. Meso1, a basic-helix-loop-helix protein involved in mammalian presomitic mesoderm development. Proc Natl Acad Sci U S A. 1995 Jun 20;92(13):5870–5874. [PMC free article] [PubMed]
  • Braun T, Bober E, Arnold HH. Inhibition of muscle differentiation by the adenovirus E1a protein: repression of the transcriptional activating function of the HLH protein Myf-5. Genes Dev. 1992 May;6(5):888–902. [PubMed]
  • Burgess R, Cserjesi P, Ligon KL, Olson EN. Paraxis: a basic helix-loop-helix protein expressed in paraxial mesoderm and developing somites. Dev Biol. 1995 Apr;168(2):296–306. [PubMed]
  • Chen C, Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. [PMC free article] [PubMed]
  • Chen ZF, Behringer RR. twist is required in head mesenchyme for cranial neural tube morphogenesis. Genes Dev. 1995 Mar 15;9(6):686–699. [PubMed]
  • Cossu G, Tajbakhsh S, Buckingham M. How is myogenesis initiated in the embryo? Trends Genet. 1996 Jun;12(6):218–223. [PubMed]
  • Dang CV, McGuire M, Buckmire M, Lee WM. Involvement of the 'leucine zipper' region in the oligomerization and transforming activity of human c-myc protein. Nature. 1989 Feb 16;337(6208):664–666. [PubMed]
  • Davis RL, Cheng PF, Lassar AB, Weintraub H. The MyoD DNA binding domain contains a recognition code for muscle-specific gene activation. Cell. 1990 Mar 9;60(5):733–746. [PubMed]
  • Davis RL, Weintraub H. Acquisition of myogenic specificity by replacement of three amino acid residues from MyoD into E12. Science. 1992 May 15;256(5059):1027–1030. [PubMed]
  • Fairman R, Beran-Steed RK, Anthony-Cahill SJ, Lear JD, Stafford WF, 3rd, DeGrado WF, Benfield PA, Brenner SL. Multiple oligomeric states regulate the DNA binding of helix-loop-helix peptides. Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10429–10433. [PMC free article] [PubMed]
  • Farmer K, Catala F, Wright WE. Alternative multimeric structures affect myogenin DNA binding activity. J Biol Chem. 1992 Mar 15;267(8):5631–5636. [PubMed]
  • Ferré-D'Amaré AR, Pognonec P, Roeder RG, Burley SK. Structure and function of the b/HLH/Z domain of USF. EMBO J. 1994 Jan 1;13(1):180–189. [PMC free article] [PubMed]
  • Fisher DE, Carr CS, Parent LA, Sharp PA. TFEB has DNA-binding and oligomerization properties of a unique helix-loop-helix/leucine-zipper family. Genes Dev. 1991 Dec;5(12A):2342–2352. [PubMed]
  • Franco del Amo F, Gendron-Maguire M, Swiatek PJ, Gridley T. Cloning, sequencing and expression of the mouse mammalian achaete-scute homolog 1 (MASH1). Biochim Biophys Acta. 1993 Jan 23;1171(3):323–327. [PubMed]
  • Füchtbauer EM. Expression of M-twist during postimplantation development of the mouse. Dev Dyn. 1995 Nov;204(3):316–322. [PubMed]
  • Göbel V, Lipkowitz S, Kozak CA, Kirsch IR. NSCL-2: a basic domain helix-loop-helix gene expressed in early neurogenesis. Cell Growth Differ. 1992 Mar;3(3):143–148. [PubMed]
  • Groisman R, Masutani H, Leibovitch MP, Robin P, Soudant I, Trouche D, Harel-Bellan A. Physical interaction between the mitogen-responsive serum response factor and myogenic basic-helix-loop-helix proteins. J Biol Chem. 1996 Mar 1;271(9):5258–5264. [PubMed]
  • Hebrok M, Wertz K, Füchtbauer EM. M-twist is an inhibitor of muscle differentiation. Dev Biol. 1994 Oct;165(2):537–544. [PubMed]
  • Hollenberg SM, Sternglanz R, Cheng PF, Weintraub H. Identification of a new family of tissue-specific basic helix-loop-helix proteins with a two-hybrid system. Mol Cell Biol. 1995 Jul;15(7):3813–3822. [PMC free article] [PubMed]
  • Hu JS, Olson EN, Kingston RE. HEB, a helix-loop-helix protein related to E2A and ITF2 that can modulate the DNA-binding ability of myogenic regulatory factors. Mol Cell Biol. 1992 Mar;12(3):1031–1042. [PMC free article] [PubMed]
  • Huang J, Blackwell TK, Kedes L, Weintraub H. Differences between MyoD DNA binding and activation site requirements revealed by functional random sequence selection. Mol Cell Biol. 1996 Jul;16(7):3893–3900. [PMC free article] [PubMed]
  • Jan YN, Jan LY. HLH proteins, fly neurogenesis, and vertebrate myogenesis. Cell. 1993 Dec 3;75(5):827–830. [PubMed]
  • Kadesch T. Consequences of heteromeric interactions among helix-loop-helix proteins. Cell Growth Differ. 1993 Jan;4(1):49–55. [PubMed]
  • Kaushal S, Schneider JW, Nadal-Ginard B, Mahdavi V. Activation of the myogenic lineage by MEF2A, a factor that induces and cooperates with MyoD. Science. 1994 Nov 18;266(5188):1236–1240. [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]
  • Lee JE, Hollenberg SM, Snider L, Turner DL, Lipnick N, Weintraub H. Conversion of Xenopus ectoderm into neurons by NeuroD, a basic helix-loop-helix protein. Science. 1995 May 12;268(5212):836–844. [PubMed]
  • Leszczynski JF, Rose GD. Loops in globular proteins: a novel category of secondary structure. Science. 1986 Nov 14;234(4778):849–855. [PubMed]
  • Li L, Cserjesi P, Olson EN. Dermo-1: a novel twist-related bHLH protein expressed in the developing dermis. Dev Biol. 1995 Nov;172(1):280–292. [PubMed]
  • Ma PC, Rould MA, Weintraub H, Pabo CO. Crystal structure of MyoD bHLH domain-DNA complex: perspectives on DNA recognition and implications for transcriptional activation. Cell. 1994 May 6;77(3):451–459. [PubMed]
  • Molkentin JD, Black BL, Martin JF, Olson EN. Cooperative activation of muscle gene expression by MEF2 and myogenic bHLH proteins. Cell. 1995 Dec 29;83(7):1125–1136. [PubMed]
  • Murre C, McCaw PS, Baltimore D. A new DNA binding and dimerization motif in immunoglobulin enhancer binding, daughterless, MyoD, and myc proteins. Cell. 1989 Mar 10;56(5):777–783. [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]
  • 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]
  • Olson EN, Perry M, Schulz RA. Regulation of muscle differentiation by the MEF2 family of MADS box transcription factors. Dev Biol. 1995 Nov;172(1):2–14. [PubMed]
  • Osada H, Grutz G, Axelson H, Forster A, Rabbitts TH. Association of erythroid transcription factors: complexes involving the LIM protein RBTN2 and the zinc-finger protein GATA1. Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9585–9589. [PMC free article] [PubMed]
  • Ott MO, Bober E, Lyons G, Arnold H, Buckingham M. Early expression of the myogenic regulatory gene, myf-5, in precursor cells of skeletal muscle in the mouse embryo. Development. 1991 Apr;111(4):1097–1107. [PubMed]
  • Peltenburg LT, Murre C. Engrailed and Hox homeodomain proteins contain a related Pbx interaction motif that recognizes a common structure present in Pbx. EMBO J. 1996 Jul 1;15(13):3385–3393. [PMC free article] [PubMed]
  • Pesce S, Benezra R. The loop region of the helix-loop-helix protein Id1 is critical for its dominant negative activity. Mol Cell Biol. 1993 Dec;13(12):7874–7880. [PMC free article] [PubMed]
  • Pownall ME, Emerson CP., Jr Sequential activation of three myogenic regulatory genes during somite morphogenesis in quail embryos. Dev Biol. 1992 May;151(1):67–79. [PubMed]
  • Quertermous EE, Hidai H, Blanar MA, Quertermous T. Cloning and characterization of a basic helix-loop-helix protein expressed in early mesoderm and the developing somites. Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7066–7070. [PMC free article] [PubMed]
  • Roberts VJ, Steenbergen R, Murre C. Localization of E2A mRNA expression in developing and adult rat tissues. Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7583–7587. [PMC free article] [PubMed]
  • Sartorelli V, Hong NA, Bishopric NH, Kedes L. Myocardial activation of the human cardiac alpha-actin promoter by helix-loop-helix proteins. Proc Natl Acad Sci U S A. 1992 May 1;89(9):4047–4051. [PMC free article] [PubMed]
  • Sartorelli V, Webster KA, Kedes L. Muscle-specific expression of the cardiac alpha-actin gene requires MyoD1, CArG-box binding factor, and Sp1. Genes Dev. 1990 Oct;4(10):1811–1822. [PubMed]
  • Sartorelli V, Huang J, Hamamori Y, Kedes L. Molecular mechanisms of myogenic coactivation by p300: direct interaction with the activation domain of MyoD and with the MADS box of MEF2C. Mol Cell Biol. 1997 Feb;17(2):1010–1026. [PMC free article] [PubMed]
  • Sassoon DA. Myogenic regulatory factors: dissecting their role and regulation during vertebrate embryogenesis. Dev Biol. 1993 Mar;156(1):11–23. [PubMed]
  • Smith TH, Kachinsky AM, Miller JB. Somite subdomains, muscle cell origins, and the four muscle regulatory factor proteins. J Cell Biol. 1994 Oct;127(1):95–105. [PMC free article] [PubMed]
  • Spicer DB, Rhee J, Cheung WL, Lassar AB. Inhibition of myogenic bHLH and MEF2 transcription factors by the bHLH protein Twist. Science. 1996 Jun 7;272(5267):1476–1480. [PubMed]
  • Stoetzel C, Weber B, Bourgeois P, Bolcato-Bellemin AL, Perrin-Schmitt F. Dorso-ventral and rostro-caudal sequential expression of M-twist in the postimplantation murine embryo. Mech Dev. 1995 Jun;51(2-3):251–263. [PubMed]
  • Tapscott SJ, Davis RL, Thayer MJ, Cheng PF, Weintraub H, Lassar AB. MyoD1: a nuclear phosphoprotein requiring a Myc homology region to convert fibroblasts to myoblasts. Science. 1988 Oct 21;242(4877):405–411. [PubMed]
  • Taylor DA, Kraus VB, Schwarz JJ, Olson EN, Kraus WE. E1A-mediated inhibition of myogenesis correlates with a direct physical interaction of E1A12S and basic helix-loop-helix proteins. Mol Cell Biol. 1993 Aug;13(8):4714–4727. [PMC free article] [PubMed]
  • Thisse C, Thisse B. Dorsoventral development of the Drosophila embryo is controlled by a cascade of transcriptional regulators. Dev Suppl. 1992:173–181. [PubMed]
  • Um M, Li C, Manley JL. The transcriptional repressor even-skipped interacts directly with TATA-binding protein. Mol Cell Biol. 1995 Sep;15(9):5007–5016. [PMC free article] [PubMed]
  • Voronova A, Baltimore D. Mutations that disrupt DNA binding and dimer formation in the E47 helix-loop-helix protein map to distinct domains. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4722–4726. [PMC free article] [PubMed]
  • Weintraub H. The MyoD family and myogenesis: redundancy, networks, and thresholds. Cell. 1993 Dec 31;75(7):1241–1244. [PubMed]
  • Weintraub H, Davis R, Lockshon D, Lassar A. MyoD binds cooperatively to two sites in a target enhancer sequence: occupancy of two sites is required for activation. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5623–5627. [PMC free article] [PubMed]
  • Weintraub H, Dwarki VJ, Verma I, Davis R, Hollenberg S, Snider L, Lassar A, Tapscott SJ. Muscle-specific transcriptional activation by MyoD. Genes Dev. 1991 Aug;5(8):1377–1386. [PubMed]
  • Weintraub H, Genetta T, Kadesch T. Tissue-specific gene activation by MyoD: determination of specificity by cis-acting repression elements. Genes Dev. 1994 Sep 15;8(18):2203–2211. [PubMed]
  • Williams JS, Andrisani OM. The hepatitis B virus X protein targets the basic region-leucine zipper domain of CREB. Proc Natl Acad Sci U S A. 1995 Apr 25;92(9):3819–3823. [PMC free article] [PubMed]
  • Wolf C, Thisse C, Stoetzel C, Thisse B, Gerlinger P, Perrin-Schmitt F. The M-twist gene of Mus is expressed in subsets of mesodermal cells and is closely related to the Xenopus X-twi and the Drosophila twist genes. Dev Biol. 1991 Feb;143(2):363–373. [PubMed]
  • Yuan W, Condorelli G, Caruso M, Felsani A, Giordano A. Human p300 protein is a coactivator for the transcription factor MyoD. J Biol Chem. 1996 Apr 12;271(15):9009–9013. [PubMed]
  • Yun K, Wold B. Skeletal muscle determination and differentiation: story of a core regulatory network and its context. Curr Opin Cell Biol. 1996 Dec;8(6):877–889. [PubMed]
  • Zhang Y, Doyle K, Bina M. Interactions of HTF4 with E-box motifs in the long terminal repeat of human immunodeficiency virus type 1. J Virol. 1992 Sep;66(9):5631–5634. [PMC free article] [PubMed]
  • Zhou Q, Gedrich RW, Engel DA. Transcriptional repression of the c-fos gene by YY1 is mediated by a direct interaction with ATF/CREB. J Virol. 1995 Jul;69(7):4323–4330. [PMC free article] [PubMed]

Articles from Molecular and Cellular Biology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...