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. 1996 Oct;16(10):5865–5875. doi: 10.1128/mcb.16.10.5865

Two new members of the murine Sim gene family are transcriptional repressors and show different expression patterns during mouse embryogenesis.

M Ema 1, M Morita 1, S Ikawa 1, M Tanaka 1, Y Matsuda 1, O Gotoh 1, Y Saijoh 1, H Fujii 1, H Hamada 1, Y Kikuchi 1, Y Fujii-Kuriyama 1
PMCID: PMC231588  PMID: 8927054

Abstract

From a cDNA library of mouse skeletal muscle, we have isolated mouse Sim1 (mSim1) cDNA encoding a polypeptide of 765 amino acids with striking amino acid identify in basic helix-loop-helix (89% identify) and PAS (89 % identify) domains to previously identified mSim2, although the carboxy-terminal third of the molecule did not show any similarity to mSim2 or Drosophila Sim (dSim). Yeast two-hybrid analysis and coimmunoprecipitation experiments demonstrated that both of the mSim gene products interacted with Arnt even more efficiently than AhR, a natural partner of Arnt, suggesting a functional cooperativity with Arnt. In sharp contrast with dSim having transcriptional-enhancing activity in the carboxy-terminal region, the two mSims possessed a repressive activity toward Arnt in the heterodimer complex. This is the first example of bHLH-PAS proteins with transrepressor activity, although some genetic data suggest that dSim plays a repressive role in gene expression (Z. Chang, D. Price, S. Bockheim, M. J. Boedigheimer, R. Smith, and A. Laughon, Dev. Biol. 160:315-322, 1993; D. M. Mellerick and M. Nirenberg, Dev. Biol. 171:306-316, 1995). Whole-mount in situ hybridization showed restricted and characteristic expression patterns of the two mSim mRNAs in various tissues and organs during embryogenesis, such as those for the somite, the nephrogenic cord, and the mesencephalon (for mSim1) and those for the diencephalon, branchial arches, and limbs (for mSim2). From sequence similarity and chromosomal localization, it is concluded that mSim2 is an ortholog of hSim2, which is proposed to be a candidate gene responsible for Down's syndrome. The sites of mSim2 expression showed an overlap with the affected regions of the syndrome, further strengthening involvement of mSim2 in Down's syndrome.

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Selected References

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  1. Ayer D. E., Kretzner L., Eisenman R. N. Mad: a heterodimeric partner for Max that antagonizes Myc transcriptional activity. Cell. 1993 Jan 29;72(2):211–222. doi: 10.1016/0092-8674(93)90661-9. [DOI] [PubMed] [Google Scholar]
  2. Ayer D. E., Lawrence Q. A., Eisenman R. N. Mad-Max transcriptional repression is mediated by ternary complex formation with mammalian homologs of yeast repressor Sin3. Cell. 1995 Mar 10;80(5):767–776. doi: 10.1016/0092-8674(95)90355-0. [DOI] [PubMed] [Google Scholar]
  3. Baeuerle P. A., Henkel T. Function and activation of NF-kappa B in the immune system. Annu Rev Immunol. 1994;12:141–179. doi: 10.1146/annurev.iy.12.040194.001041. [DOI] [PubMed] [Google Scholar]
  4. Burbach K. M., Poland A., Bradfield C. A. Cloning of the Ah-receptor cDNA reveals a distinctive ligand-activated transcription factor. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):8185–8189. doi: 10.1073/pnas.89.17.8185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chang Z., Price B. D., Bockheim S., Boedigheimer M. J., Smith R., Laughon A. Molecular and genetic characterization of the Drosophila tartan gene. Dev Biol. 1993 Dec;160(2):315–332. doi: 10.1006/dbio.1993.1310. [DOI] [PubMed] [Google Scholar]
  6. Chen H., Chrast R., Rossier C., Gos A., Antonarakis S. E., Kudoh J., Yamaki A., Shindoh N., Maeda H., Minoshima S. Single-minded and Down syndrome? Nat Genet. 1995 May;10(1):9–10. doi: 10.1038/ng0595-9. [DOI] [PubMed] [Google Scholar]
  7. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  8. Crews S. T., Thomas J. B., Goodman C. S. The Drosophila single-minded gene encodes a nuclear protein with sequence similarity to the per gene product. Cell. 1988 Jan 15;52(1):143–151. doi: 10.1016/0092-8674(88)90538-7. [DOI] [PubMed] [Google Scholar]
  9. Dahmane N., Charron G., Lopes C., Yaspo M. L., Maunoury C., Decorte L., Sinet P. M., Bloch B., Delabar J. M. Down syndrome-critical region contains a gene homologous to Drosophila sim expressed during rat and human central nervous system development. Proc Natl Acad Sci U S A. 1995 Sep 26;92(20):9191–9195. doi: 10.1073/pnas.92.20.9191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Echelard Y., Epstein D. J., St-Jacques B., Shen L., Mohler J., McMahon J. A., McMahon A. P. Sonic hedgehog, a member of a family of putative signaling molecules, is implicated in the regulation of CNS polarity. Cell. 1993 Dec 31;75(7):1417–1430. doi: 10.1016/0092-8674(93)90627-3. [DOI] [PubMed] [Google Scholar]
  11. Ema M., Ohe N., Suzuki M., Mimura J., Sogawa K., Ikawa S., Fujii-Kuriyama Y. Dioxin binding activities of polymorphic forms of mouse and human arylhydrocarbon receptors. J Biol Chem. 1994 Nov 4;269(44):27337–27343. [PubMed] [Google Scholar]
  12. Ema M., Sogawa K., Watanabe N., Chujoh Y., Matsushita N., Gotoh O., Funae Y., Fujii-Kuriyama Y. cDNA cloning and structure of mouse putative Ah receptor. Biochem Biophys Res Commun. 1992 Apr 15;184(1):246–253. doi: 10.1016/0006-291x(92)91185-s. [DOI] [PubMed] [Google Scholar]
  13. Ema M., Suzuki M., Morita M., Hirose K., Sogawa K., Matsuda Y., Gotoh O., Saijoh Y., Fujii H., Hamada H. cDNA cloning of a murine homologue of Drosophila single-minded, its mRNA expression in mouse development, and chromosome localization. Biochem Biophys Res Commun. 1996 Jan 17;218(2):588–594. doi: 10.1006/bbrc.1996.0104. [DOI] [PubMed] [Google Scholar]
  14. Ericson J., Muhr J., Placzek M., Lints T., Jessell T. M., Edlund T. Sonic hedgehog induces the differentiation of ventral forebrain neurons: a common signal for ventral patterning within the neural tube. Cell. 1995 Jun 2;81(5):747–756. doi: 10.1016/0092-8674(95)90536-7. [DOI] [PubMed] [Google Scholar]
  15. Fan C. M., Kuwana E., Bulfone A., Fletcher C. F., Copeland N. G., Jenkins N. A., Crews S., Martinez S., Puelles L., Rubenstein J. L. Expression patterns of two murine homologs of Drosophila single-minded suggest possible roles in embryonic patterning and in the pathogenesis of Down syndrome. Mol Cell Neurosci. 1996 Jan;7(1):1–16. doi: 10.1006/mcne.1996.0001. [DOI] [PubMed] [Google Scholar]
  16. Franks R. G., Crews S. T. Transcriptional activation domains of the single-minded bHLH protein are required for CNS midline cell development. Mech Dev. 1994 Mar;45(3):269–277. doi: 10.1016/0925-4773(94)90013-2. [DOI] [PubMed] [Google Scholar]
  17. Gashler A. L., Swaminathan S., Sukhatme V. P. A novel repression module, an extensive activation domain, and a bipartite nuclear localization signal defined in the immediate-early transcription factor Egr-1. Mol Cell Biol. 1993 Aug;13(8):4556–4571. doi: 10.1128/mcb.13.8.4556. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gotoh O. A weighting system and algorithm for aligning many phylogenetically related sequences. Comput Appl Biosci. 1995 Oct;11(5):543–551. doi: 10.1093/bioinformatics/11.5.543. [DOI] [PubMed] [Google Scholar]
  20. Han K., Manley J. L. Transcriptional repression by the Drosophila even-skipped protein: definition of a minimal repression domain. Genes Dev. 1993 Mar;7(3):491–503. doi: 10.1101/gad.7.3.491. [DOI] [PubMed] [Google Scholar]
  21. Hirose K., Morita M., Ema M., Mimura J., Hamada H., Fujii H., Saijo Y., Gotoh O., Sogawa K., Fujii-Kuriyama Y. cDNA cloning and tissue-specific expression of a novel basic helix-loop-helix/PAS factor (Arnt2) with close sequence similarity to the aryl hydrocarbon receptor nuclear translocator (Arnt). Mol Cell Biol. 1996 Apr;16(4):1706–1713. doi: 10.1128/mcb.16.4.1706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hoffman E. C., Reyes H., Chu F. F., Sander F., Conley L. H., Brooks B. A., Hankinson O. Cloning of a factor required for activity of the Ah (dioxin) receptor. Science. 1991 May 17;252(5008):954–958. doi: 10.1126/science.1852076. [DOI] [PubMed] [Google Scholar]
  23. Hynes M., Poulsen K., Tessier-Lavigne M., Rosenthal A. Control of neuronal diversity by the floor plate: contact-mediated induction of midbrain dopaminergic neurons. Cell. 1995 Jan 13;80(1):95–101. doi: 10.1016/0092-8674(95)90454-9. [DOI] [PubMed] [Google Scholar]
  24. Isaac D. D., Andrew D. J. Tubulogenesis in Drosophila: a requirement for the trachealess gene product. Genes Dev. 1996 Jan 1;10(1):103–117. doi: 10.1101/gad.10.1.103. [DOI] [PubMed] [Google Scholar]
  25. Jackson F. R., Bargiello T. A., Yun S. H., Young M. W. Product of per locus of Drosophila shares homology with proteoglycans. Nature. 1986 Mar 13;320(6058):185–188. doi: 10.1038/320185a0. [DOI] [PubMed] [Google Scholar]
  26. Jain S., Dolwick K. M., Schmidt J. V., Bradfield C. A. Potent transactivation domains of the Ah receptor and the Ah receptor nuclear translocator map to their carboxyl termini. J Biol Chem. 1994 Dec 16;269(50):31518–31524. [PubMed] [Google Scholar]
  27. Klämbt C., Jacobs J. R., Goodman C. S. The midline of the Drosophila central nervous system: a model for the genetic analysis of cell fate, cell migration, and growth cone guidance. Cell. 1991 Feb 22;64(4):801–815. doi: 10.1016/0092-8674(91)90509-w. [DOI] [PubMed] [Google Scholar]
  28. Kozak M. The scanning model for translation: an update. J Cell Biol. 1989 Feb;108(2):229–241. doi: 10.1083/jcb.108.2.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Krauss S., Concordet J. P., Ingham P. W. A functionally conserved homolog of the Drosophila segment polarity gene hh is expressed in tissues with polarizing activity in zebrafish embryos. Cell. 1993 Dec 31;75(7):1431–1444. doi: 10.1016/0092-8674(93)90628-4. [DOI] [PubMed] [Google Scholar]
  30. Li H., Dong L., Whitlock J. P., Jr Transcriptional activation function of the mouse Ah receptor nuclear translocator. J Biol Chem. 1994 Nov 11;269(45):28098–28105. [PubMed] [Google Scholar]
  31. Licht J. D., Grossel M. J., Figge J., Hansen U. M. Drosophila Krüppel protein is a transcriptional repressor. Nature. 1990 Jul 5;346(6279):76–79. doi: 10.1038/346076a0. [DOI] [PubMed] [Google Scholar]
  32. Lillie J. W., Green M. R. Transcription activation by the adenovirus E1a protein. Nature. 1989 Mar 2;338(6210):39–44. doi: 10.1038/338039a0. [DOI] [PubMed] [Google Scholar]
  33. Lindebro M. C., Poellinger L., Whitelaw M. L. Protein-protein interaction via PAS domains: role of the PAS domain in positive and negative regulation of the bHLH/PAS dioxin receptor-Arnt transcription factor complex. EMBO J. 1995 Jul 17;14(14):3528–3539. doi: 10.1002/j.1460-2075.1995.tb07359.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Madden S. L., Cook D. M., Morris J. F., Gashler A., Sukhatme V. P., Rauscher F. J., 3rd Transcriptional repression mediated by the WT1 Wilms tumor gene product. Science. 1991 Sep 27;253(5027):1550–1553. doi: 10.1126/science.1654597. [DOI] [PubMed] [Google Scholar]
  35. Madden S. L., Cook D. M., Rauscher F. J., 3rd A structure-function analysis of transcriptional repression mediated by the WT1, Wilms' tumor suppressor protein. Oncogene. 1993 Jul;8(7):1713–1720. [PubMed] [Google Scholar]
  36. Matsuda Y., Chapman V. M. Application of fluorescence in situ hybridization in genome analysis of the mouse. Electrophoresis. 1995 Feb;16(2):261–272. doi: 10.1002/elps.1150160142. [DOI] [PubMed] [Google Scholar]
  37. Matsuda Y., Harada Y. N., Natsuume-Sakai S., Lee K., Shiomi T., Chapman V. M. Location of the mouse complement factor H gene (cfh) by FISH analysis and replication R-banding. Cytogenet Cell Genet. 1992;61(4):282–285. doi: 10.1159/000133423. [DOI] [PubMed] [Google Scholar]
  38. Matsushita N., Sogawa K., Ema M., Yoshida A., Fujii-Kuriyama Y. A factor binding to the xenobiotic responsive element (XRE) of P-4501A1 gene consists of at least two helix-loop-helix proteins, Ah receptor and Arnt. J Biol Chem. 1993 Oct 5;268(28):21002–21006. [PubMed] [Google Scholar]
  39. Mellerick D. M., Nirenberg M. Dorsal-ventral patterning genes restrict NK-2 homeobox gene expression to the ventral half of the central nervous system of Drosophila embryos. Dev Biol. 1995 Oct;171(2):306–316. doi: 10.1006/dbio.1995.1283. [DOI] [PubMed] [Google Scholar]
  40. Mizushima S., Nagata S. pEF-BOS, a powerful mammalian expression vector. Nucleic Acids Res. 1990 Sep 11;18(17):5322–5322. doi: 10.1093/nar/18.17.5322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Muenke M., Bone L. J., Mitchell H. F., Hart I., Walton K., Hall-Johnson K., Ippel E. F., Dietz-Band J., Kvaløy K., Fan C. M. Physical mapping of the holoprosencephaly critical region in 21q22.3, exclusion of SIM2 as a candidate gene for holoprosencephaly, and mapping of SIM2 to a region of chromosome 21 important for Down syndrome. Am J Hum Genet. 1995 Nov;57(5):1074–1079. [PMC free article] [PubMed] [Google Scholar]
  42. Nambu J. R., Franks R. G., Hu S., Crews S. T. The single-minded gene of Drosophila is required for the expression of genes important for the development of CNS midline cells. Cell. 1990 Oct 5;63(1):63–75. doi: 10.1016/0092-8674(90)90288-p. [DOI] [PubMed] [Google Scholar]
  43. Nambu J. R., Lewis J. O., Crews S. T. The development and function of the Drosophila CNS midline cells. Comp Biochem Physiol Comp Physiol. 1993 Mar;104(3):399–409. doi: 10.1016/0300-9629(93)90439-b. [DOI] [PubMed] [Google Scholar]
  44. Nambu J. R., Lewis J. O., Wharton K. A., Jr, Crews S. T. The Drosophila single-minded gene encodes a helix-loop-helix protein that acts as a master regulator of CNS midline development. Cell. 1991 Dec 20;67(6):1157–1167. doi: 10.1016/0092-8674(91)90292-7. [DOI] [PubMed] [Google Scholar]
  45. Pourquié O., Fan C. M., Coltey M., Hirsinger E., Watanabe Y., Bréant C., Francis-West P., Brickell P., Tessier-Lavigne M., Le Douarin N. M. Lateral and axial signals involved in avian somite patterning: a role for BMP4. Cell. 1996 Feb 9;84(3):461–471. doi: 10.1016/s0092-8674(00)81291-x. [DOI] [PubMed] [Google Scholar]
  46. Reddy P., Jacquier A. C., Abovich N., Petersen G., Rosbash M. The period clock locus of D. melanogaster codes for a proteoglycan. Cell. 1986 Jul 4;46(1):53–61. doi: 10.1016/0092-8674(86)90859-7. [DOI] [PubMed] [Google Scholar]
  47. Reyes H., Reisz-Porszasz S., Hankinson O. Identification of the Ah receptor nuclear translocator protein (Arnt) as a component of the DNA binding form of the Ah receptor. Science. 1992 May 22;256(5060):1193–1195. doi: 10.1126/science.256.5060.1193. [DOI] [PubMed] [Google Scholar]
  48. Riddle R. D., Johnson R. L., Laufer E., Tabin C. Sonic hedgehog mediates the polarizing activity of the ZPA. Cell. 1993 Dec 31;75(7):1401–1416. doi: 10.1016/0092-8674(93)90626-2. [DOI] [PubMed] [Google Scholar]
  49. Roelink H., Augsburger A., Heemskerk J., Korzh V., Norlin S., Ruiz i Altaba A., Tanabe Y., Placzek M., Edlund T., Jessell T. M. Floor plate and motor neuron induction by vhh-1, a vertebrate homolog of hedgehog expressed by the notochord. Cell. 1994 Feb 25;76(4):761–775. doi: 10.1016/0092-8674(94)90514-2. [DOI] [PubMed] [Google Scholar]
  50. Rothberg J. M., Jacobs J. R., Goodman C. S., Artavanis-Tsakonas S. slit: an extracellular protein necessary for development of midline glia and commissural axon pathways contains both EGF and LRR domains. Genes Dev. 1990 Dec;4(12A):2169–2187. doi: 10.1101/gad.4.12a.2169. [DOI] [PubMed] [Google Scholar]
  51. Rubenstein J. L., Martinez S., Shimamura K., Puelles L. The embryonic vertebrate forebrain: the prosomeric model. Science. 1994 Oct 28;266(5185):578–580. doi: 10.1126/science.7939711. [DOI] [PubMed] [Google Scholar]
  52. Sadowski I., Ptashne M. A vector for expressing GAL4(1-147) fusions in mammalian cells. Nucleic Acids Res. 1989 Sep 25;17(18):7539–7539. doi: 10.1093/nar/17.18.7539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Saitou N., Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987 Jul;4(4):406–425. doi: 10.1093/oxfordjournals.molbev.a040454. [DOI] [PubMed] [Google Scholar]
  54. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Shi Y., Seto E., Chang L. S., Shenk T. Transcriptional repression by YY1, a human GLI-Krüppel-related protein, and relief of repression by adenovirus E1A protein. Cell. 1991 Oct 18;67(2):377–388. doi: 10.1016/0092-8674(91)90189-6. [DOI] [PubMed] [Google Scholar]
  56. Sogawa K., Iwabuchi K., Abe H., Fujii-Kuriyama Y. Transcriptional activation domains of the Ah receptor and Ah receptor nuclear translocator. J Cancer Res Clin Oncol. 1995;121(9-10):612–620. doi: 10.1007/BF01197779. [DOI] [PubMed] [Google Scholar]
  57. Sogawa K., Nakano R., Kobayashi A., Kikuchi Y., Ohe N., Matsushita N., Fujii-Kuriyama Y. Possible function of Ah receptor nuclear translocator (Arnt) homodimer in transcriptional regulation. Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):1936–1940. doi: 10.1073/pnas.92.6.1936. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Takahashi J. S. Circadian clock genes are ticking. Science. 1992 Oct 9;258(5080):238–240. doi: 10.1126/science.1384127. [DOI] [PubMed] [Google Scholar]
  59. Thomas J. B., Crews S. T., Goodman C. S. Molecular genetics of the single-minded locus: a gene involved in the development of the Drosophila nervous system. Cell. 1988 Jan 15;52(1):133–141. doi: 10.1016/0092-8674(88)90537-5. [DOI] [PubMed] [Google Scholar]
  60. Wang G. L., Jiang B. H., Rue E. A., Semenza G. L. Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension. Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5510–5514. doi: 10.1073/pnas.92.12.5510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Weintraub H., Davis R., Tapscott S., Thayer M., Krause M., Benezra R., Blackwell T. K., Turner D., Rupp R., Hollenberg S. The myoD gene family: nodal point during specification of the muscle cell lineage. Science. 1991 Feb 15;251(4995):761–766. doi: 10.1126/science.1846704. [DOI] [PubMed] [Google Scholar]
  62. Wharton K. A., Jr, Franks R. G., Kasai Y., Crews S. T. Control of CNS midline transcription by asymmetric E-box-like elements: similarity to xenobiotic responsive regulation. Development. 1994 Dec;120(12):3563–3569. doi: 10.1242/dev.120.12.3563. [DOI] [PubMed] [Google Scholar]
  63. Whitelaw M. L., Gustafsson J. A., Poellinger L. Identification of transactivation and repression functions of the dioxin receptor and its basic helix-loop-helix/PAS partner factor Arnt: inducible versus constitutive modes of regulation. Mol Cell Biol. 1994 Dec;14(12):8343–8355. doi: 10.1128/mcb.14.12.8343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. Whitelaw M., Pongratz I., Wilhelmsson A., Gustafsson J. A., Poellinger L. Ligand-dependent recruitment of the Arnt coregulator determines DNA recognition by the dioxin receptor. Mol Cell Biol. 1993 Apr;13(4):2504–2514. doi: 10.1128/mcb.13.4.2504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. Wilk R., Weizman I., Shilo B. Z. trachealess encodes a bHLH-PAS protein that is an inducer of tracheal cell fates in Drosophila. Genes Dev. 1996 Jan 1;10(1):93–102. doi: 10.1101/gad.10.1.93. [DOI] [PubMed] [Google Scholar]
  66. Zervos A. S., Gyuris J., Brent R. Mxi1, a protein that specifically interacts with Max to bind Myc-Max recognition sites. Cell. 1993 Jan 29;72(2):223–232. doi: 10.1016/0092-8674(93)90662-a. [DOI] [PubMed] [Google Scholar]

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