PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of molcellbPermissionsJournals.ASM.orgJournalMCB ArticleJournal InfoAuthorsReviewers
 
Mol Cell Biol. 1997 May; 17(5): 2920–2932.
PMCID: PMC232144

Heterodimerization of Msx and Dlx homeoproteins results in functional antagonism.

Abstract

Protein-protein interactions are known to be essential for specifying the transcriptional activities of homeoproteins. Here we show that representative members of the Msx and Dlx homeoprotein families form homo- and heterodimeric complexes. We demonstrate that dimerization by Msx and Dlx proteins is mediated through their homeodomains and that the residues required for this interaction correspond to those necessary for DNA binding. Unlike most other known examples of homeoprotein interactions, association of Msx and Dlx proteins does not promote cooperative DNA binding; instead, dimerization and DNA binding are mutually exclusive activities. In particular, we show that Msx and Dlx proteins interact independently and noncooperatively with homeodomain DNA binding sites and that dimerization is specifically blocked by the presence of such DNA sites. We further demonstrate that the transcriptional properties of Msx and Dlx proteins display reciprocal inhibition. Specifically, Msx proteins act as transcriptional repressors and Dlx proteins act as activators, while in combination, Msx and Dlx proteins counteract each other's transcriptional activities. Finally, we show that the expression patterns of representative Msx and Dlx genes (Msx1, Msx2, Dlx2, and Dlx5) overlap in mouse embryogenesis during limb bud and craniofacial development, consistent with the potential for their protein products to interact in vivo. Based on these observations, we propose that functional antagonism through heterodimer formation provides a mechanism for regulating the transcriptional actions of Msx and Dlx homeoproteins in vivo.

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.
  • Brown JM, Wedden SE, Millburn GH, Robson LG, Hill RE, Davidson DR, Tickle C. Experimental analysis of the control of expression of the homeobox-gene Msx-1 in the developing limb and face. Development. 1993 Sep;119(1):41–48. [PubMed]
  • Catron KM, Iler N, Abate C. Nucleotides flanking a conserved TAAT core dictate the DNA binding specificity of three murine homeodomain proteins. Mol Cell Biol. 1993 Apr;13(4):2354–2365. [PMC free article] [PubMed]
  • Catron KM, Wang H, Hu G, Shen MM, Abate-Shen C. Comparison of MSX-1 and MSX-2 suggests a molecular basis for functional redundancy. Mech Dev. 1996 Apr;55(2):185–199. [PubMed]
  • Catron KM, Zhang H, Marshall SC, Inostroza JA, Wilson JM, Abate C. Transcriptional repression by Msx-1 does not require homeodomain DNA-binding sites. Mol Cell Biol. 1995 Feb;15(2):861–871. [PMC free article] [PubMed]
  • Chan-Thomas PS, Thompson RP, Robert B, Yacoub MH, Barton PJ. Expression of homeobox genes Msx-1 (Hox-7) and Msx-2 (Hox-8) during cardiac development in the chick. Dev Dyn. 1993 Jul;197(3):203–216. [PubMed]
  • Cleary MA, Stern S, Tanaka M, Herr W. Differential positive control by Oct-1 and Oct-2: activation of a transcriptionally silent motif through Oct-1 and VP16 corecruitment. Genes Dev. 1993 Jan;7(1):72–83. [PubMed]
  • Coelho CN, Sumoy L, Rodgers BJ, Davidson DR, Hill RE, Upholt WB, Kosher RA. Expression of the chicken homeobox-containing gene GHox-8 during embryonic chick limb development. Mech Dev. 1991 Jun;34(2-3):143–154. [PubMed]
  • Davidson DR, Crawley A, Hill RE, Tickle C. Position-dependent expression of two related homeobox genes in developing vertebrate limbs. Nature. 1991 Aug 1;352(6334):429–431. [PubMed]
  • Dollé P, Price M, Duboule D. Expression of the murine Dlx-1 homeobox gene during facial, ocular and limb development. Differentiation. 1992 Mar;49(2):93–99. [PubMed]
  • Ekker M, Akimenko MA, Bremiller R, Westerfield M. Regional expression of three homeobox transcripts in the inner ear of zebrafish embryos. Neuron. 1992 Jul;9(1):27–35. [PubMed]
  • Ferrari D, Sumoy L, Gannon J, Sun H, Brown AM, Upholt WB, Kosher RA. The expression pattern of the Distal-less homeobox-containing gene Dlx-5 in the developing chick limb bud suggests its involvement in apical ectodermal ridge activity, pattern formation, and cartilage differentiation. Mech Dev. 1995 Aug;52(2-3):257–264. [PubMed]
  • Hill RE, Jones PF, Rees AR, Sime CM, Justice MJ, Copeland NG, Jenkins NA, Graham E, Davidson DR. A new family of mouse homeo box-containing genes: molecular structure, chromosomal location, and developmental expression of Hox-7.1. Genes Dev. 1989 Jan;3(1):26–37. [PubMed]
  • Holland PW. Cloning and evolutionary analysis of msh-like homeobox genes from mouse, zebrafish and ascidian. Gene. 1991 Feb 15;98(2):253–257. [PubMed]
  • Huang CC, Herr W. Differential control of transcription by homologous homeodomain coregulators. Mol Cell Biol. 1996 Jun;16(6):2967–2976. [PMC free article] [PubMed]
  • Iler N, Rowitch DH, Echelard Y, McMahon AP, Abate-Shen C. A single homeodomain binding site restricts spatial expression of Wnt-1 in the developing brain. Mech Dev. 1995 Sep;53(1):87–96. [PubMed]
  • Jowett AK, Vainio S, Ferguson MW, Sharpe PT, Thesleff I. Epithelial-mesenchymal interactions are required for msx 1 and msx 2 gene expression in the developing murine molar tooth. Development. 1993 Feb;117(2):461–470. [PubMed]
  • Lai JS, Cleary MA, Herr W. A single amino acid exchange transfers VP16-induced positive control from the Oct-1 to the Oct-2 homeo domain. Genes Dev. 1992 Nov;6(11):2058–2065. [PubMed]
  • Lai JS, Herr W. Ethidium bromide provides a simple tool for identifying genuine DNA-independent protein associations. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):6958–6962. [PubMed]
  • Lichtsteiner S, Tjian R. Synergistic activation of transcription by UNC-86 and MEC-3 in Caenorhabditis elegans embryo extracts. EMBO J. 1995 Aug 15;14(16):3937–3945. [PubMed]
  • Lyons GE, Houzelstein D, Sassoon D, Robert B, Buckingham ME. Multiple sites of Hox-7 expression during mouse embryogenesis: comparison with retinoic acid receptor mRNA localization. Mol Reprod Dev. 1992 Aug;32(4):303–314. [PubMed]
  • MacKenzie A, Ferguson MW, Sharpe PT. Hox-7 expression during murine craniofacial development. Development. 1991 Oct;113(2):601–611. [PubMed]
  • Mann RS, Chan SK. Extra specificity from extradenticle: the partnership between HOX and PBX/EXD homeodomain proteins. Trends Genet. 1996 Jul;12(7):258–262. [PubMed]
  • Monaghan AP, Davidson DR, Sime C, Graham E, Baldock R, Bhattacharya SS, Hill RE. The Msh-like homeobox genes define domains in the developing vertebrate eye. Development. 1991 Aug;112(4):1053–1061. [PubMed]
  • Pavlova A, Boutin E, Cunha G, Sassoon D. Msx1 (Hox-7.1) in the adult mouse uterus: cellular interactions underlying regulation of expression. Development. 1994 Feb;120(2):335–345. [PubMed]
  • Porteus MH, Bulfone A, Ciaranello RD, Rubenstein JL. Isolation and characterization of a novel cDNA clone encoding a homeodomain that is developmentally regulated in the ventral forebrain. Neuron. 1991 Aug;7(2):221–229. [PubMed]
  • Qiu M, Bulfone A, Martinez S, Meneses JJ, Shimamura K, Pedersen RA, Rubenstein JL. Null mutation of Dlx-2 results in abnormal morphogenesis of proximal first and second branchial arch derivatives and abnormal differentiation in the forebrain. Genes Dev. 1995 Oct 15;9(20):2523–2538. [PubMed]
  • Reginelli AD, Wang YQ, Sassoon D, Muneoka K. Digit tip regeneration correlates with regions of Msx1 (Hox 7) expression in fetal and newborn mice. Development. 1995 Apr;121(4):1065–1076. [PubMed]
  • Robert B, Lyons G, Simandl BK, Kuroiwa A, Buckingham M. The apical ectodermal ridge regulates Hox-7 and Hox-8 gene expression in developing chick limb buds. Genes Dev. 1991 Dec;5(12B):2363–2374. [PubMed]
  • Robert B, Sassoon D, Jacq B, Gehring W, Buckingham M. Hox-7, a mouse homeobox gene with a novel pattern of expression during embryogenesis. EMBO J. 1989 Jan;8(1):91–100. [PubMed]
  • Satokata I, Maas R. Msx1 deficient mice exhibit cleft palate and abnormalities of craniofacial and tooth development. Nat Genet. 1994 Apr;6(4):348–356. [PubMed]
  • Shimeld SM, McKay IJ, Sharpe PT. The murine homeobox gene Msx-3 shows highly restricted expression in the developing neural tube. Mech Dev. 1996 Apr;55(2):201–210. [PubMed]
  • Simon HG, Nelson C, Goff D, Laufer E, Morgan BA, Tabin C. Differential expression of myogenic regulatory genes and Msx-1 during dedifferentiation and redifferentiation of regenerating amphibian limbs. Dev Dyn. 1995 Jan;202(1):1–12. [PubMed]
  • Song K, Wang Y, Sassoon D. Expression of Hox-7.1 in myoblasts inhibits terminal differentiation and induces cell transformation. Nature. 1992 Dec 3;360(6403):477–481. [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]
  • Taggart AK, Pugh BF. Dimerization of TFIID when not bound to DNA. Science. 1996 May 31;272(5266):1331–1333. [PubMed]
  • Wang Y, Sassoon D. Ectoderm-mesenchyme and mesenchyme-mesenchyme interactions regulate Msx-1 expression and cellular differentiation in the murine limb bud. Dev Biol. 1995 Apr;168(2):374–382. [PubMed]
  • Wilson D, Sheng G, Lecuit T, Dostatni N, Desplan C. Cooperative dimerization of paired class homeo domains on DNA. Genes Dev. 1993 Nov;7(11):2120–2134. [PubMed]
  • Woloshin P, Song K, Degnin C, Killary AM, Goldhamer DJ, Sassoon D, Thayer MJ. MSX1 inhibits myoD expression in fibroblast x 10T1/2 cell hybrids. Cell. 1995 Aug 25;82(4):611–620. [PubMed]
  • Xue D, Tu Y, Chalfie M. Cooperative interactions between the Caenorhabditis elegans homeoproteins UNC-86 and MEC-3. Science. 1993 Sep 3;261(5126):1324–1328. [PubMed]
  • Zappavigna V, Sartori D, Mavilio F. Specificity of HOX protein function depends on DNA-protein and protein-protein interactions, both mediated by the homeo domain. Genes Dev. 1994 Mar 15;8(6):732–744. [PubMed]
  • Zhang H, Catron KM, Abate-Shen C. A role for the Msx-1 homeodomain in transcriptional regulation: residues in the N-terminal arm mediate TATA binding protein interaction and transcriptional repression. Proc Natl Acad Sci U S A. 1996 Mar 5;93(5):1764–1769. [PubMed]

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