Overexpression of transcription factor BACH1 in fetal Down Syndrome brain

  • R. Ferrando-Miguel
  • M. S. Cheon
  • J.-W. Yang
  • G. Lubec
Part of the Journal of Neural Transmission Supplement 67 book series (NEURAL SUPPL, volume 67)


There is a series of about 12 transcription factors expressed on chromosome 21. These transcription factors (TFs) are major candidates for playing a pathogenetic role for the abnormal wiring of the brain in fetal Down Syndrome (DS) as approximately 5,000 TFs are developmentally involved in the complex architecture of the human brain. TF derangement in DS has been already reported and we decided to contribute to the problem by studying four TFs encoded on chromosome 21 in fetal DS brain.

We used fetal cortex of 8 DS fetuses and 6 controls (females) from the 18–19th week of gestation. Brain homogenates were subject to immunoblotting using goat-anti-BACH1, rabbit anti-heme oxygenase 1 (HO1), rabbit anti-ERG, rabbit anti-RUNX1 and goat anti-SIM2 l. Antibodies against beta-actin were used to normalise cell loss and antibodies against neuron-specific enolase were used to compensate neuronal loss.

BACH1 was significantly overexpressed in fetal DS (p < 0.008) as compared to controls whereas RUNX1 and ERG proteins were comparable between groups, and SIM2 1 was not detectable in any specimen. BACH1 was even significantly increased in the DS panel when normalised versus the housekeeping protein beta-actin (p < 0.01) or the neuron specific enolase (p < 0.01). HO-1 was found comparable between groups.

BACH1, a member of the family of BTB-basic leucine zipper transcription factors, regulates gene expression through the NF-E2 site. More specifically, BACH1 suppresses expression of HO1. Increased BACH1, however, did not lead to decreased HO1, which would have explained oxidative stress observed in fetal DS.


Bacterial Artificial Chromosome Down Syndrome Fetal Brain Neuron Specific Enolase Gene Dosage Effect 



Down Syndrome




Transcription regulation protein BTB and CNC homolog 1


heme oxygenase 1


Single-minded homolog 2


Runt-related transcription factor 1


Neuron specific enolase


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Asou N (2003) The role of Runt domain transcription factor AML1/RUNX1 in leukemogenesis and its clinical implications. Crit Rev Oncol/Hematol 45: 129–150CrossRefGoogle Scholar
  2. Aziz-Aloya RB, Levanon D, Kam H, Kidron D, Goldenberg D, Lotem J, Polak-Chaklon S, Groner Y (1998) Expression of AML1-d, a short human AML1 isoform, in embryonic stem cells suppress in vivo tumor growth and differentation. Cell Death Differ 5: 765–773PubMedCrossRefGoogle Scholar
  3. Bahn S, Mimmack M, Ryan M, Caldwell MA, Jauniaux E, Starkey M, Svendsen CN, Emson P (2002) Neuronal target genes of the neuron-restrictive silencer factor in neurospheres derived from fetuses with Down’s syndrome: a gene expression study. Lancet 359: 301–315CrossRefGoogle Scholar
  4. Baranano DE, Snyder SH (2001) Neural roles for heme oxygenase: contrast to nitric oxide synthase. PNAS 98: 10996–11002PubMedCrossRefGoogle Scholar
  5. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254PubMedCrossRefGoogle Scholar
  6. Busciglio J, Yanker BA (1995) Apoptosis and increased generation of reactive oxygen species in Down’s syndrome neurons in vivo. Nature 378 (6559): 776–779PubMedCrossRefGoogle Scholar
  7. Capone GT (2001) Down Syndrome: advances in molecular biology and the neurosciences. Dev Behav Pediatr 22: 40–59CrossRefGoogle Scholar
  8. Chen K, Gunter K, Maines MD (2000) Neurons overexpressing heme oxygenase-1 resist ocidative stress-mediated cell death. J Neurochem 75: 304–313PubMedCrossRefGoogle Scholar
  9. Cheon MS, Kim SH, Yaspo ML, Blasi F, Aoki Y, Melen K, Lubec G (2003) Protein levels of genes encoded on chromosome 21 in fetal Down syndrome brain: challenging the gene dosage effect hypothesis, part I. Amino Acids 24: 111–117PubMedGoogle Scholar
  10. Chrast R, Scott HS, Chen H, Kudoh J, Rossier C, Minoshima S, Wang Y, Shimizu N, Antonarakis SE (1997) Cloning of two human homologs of the Drosophila single-minded gene SIM1 on chromosome 6q and SIM2 on 21q within the Down Syndrome Chromosomal Region. Genome Res 7: 615–624PubMedGoogle Scholar
  11. Chrast R, Scott HS, Madani R, Huber L, Wolfer DP, Prinz M, Aguzzi A, Lipp HP, Antonarakis SE (2000) Mice trisomic for bacterial artificial chromosome with the single-minded 2 gene (Sim2) show phenotypes similar to some of those present in the partial trisomy 16 mouse models of Down Syndrome. Hum Mol Gen 9: 1853–1864PubMedCrossRefGoogle Scholar
  12. Dahmane N, Charron G, Lopes C, Yaspo ML, Maunoury C, Decorte L, Sinet PM, Bloch B, Delabar JM (1995) Down syndrome-critical region contains a gene homologous to Drosophila sim expressed during rat and human central nervous system development. Proc Natl Acad Sci USA 92: 9191–9195PubMedCrossRefGoogle Scholar
  13. Delattre O, Zucman J, Melot T, Garau XS, Zucker JM, Lenoir GM, Ambros PF, Sheer D, Turc-Carel C, Triche TJ, Aurias A, Thomas G (1994) The Ewing family of tumors: a subgroup of small-round-cell tumors defined by specific chimeric transcripts. N Engl J Med 331: 294–299PubMedCrossRefGoogle Scholar
  14. Dhordain P, Dewitte F, Desbiens X, Stehelin D, Duterque-Coquillaud M (1995) Mesodermal expression of the chicken erg gene associated with precartilaginous condensation and cartilage differentiation. Mech Dev 50: 17–28PubMedCrossRefGoogle Scholar
  15. Erna M, Ikegami S, Hosoya T, Mimura J, Ohtani H, Nakao K, Inokuchi K, Katasuki M, Fuji-Kuriyama Y (1999) Mild impairment of learning and memory in mice overexpressing the mSim2 gene located on chromosome 16: an animal model of Down’s syndrome. Hum Mol Gen 8: 1409–1415CrossRefGoogle Scholar
  16. Engidawork E, Lubec G (2003) Molecular changes in fetal Down Syndrome brain. J Neurochem 84: 895–904PubMedCrossRefGoogle Scholar
  17. Engidawork E, Balic N, Fountoulakis M, Dierssen M, Greber-Platzer S, Lubec G (2001) 13-Amyloid precursor protein, ETS-2 and collagen alpha 1 (VI) chain precursor, encoded in chromosome 21, are not overexpressed in fetal Down syndrome: further evidence against gene dosage effect. J Neural Transm [Suppl] 61: 335–346Google Scholar
  18. Epstein CJ (1995) Down Syndrome. In: Scriver CR, Beaudet AL, Sly WS, Valle D (eds) The metabolic and molecular bases of inherited diseade, 7th edn, vol 1. McGraw Hill, New York, pp 749–794Google Scholar
  19. Erickson P, Gao J, Chang KS, Look T, Whisenant E, Raimondi S, Lasher R, Trujillo J, Rowely J, Drabkin H (1992) Identification of breakpoints in t(8;21) acute myelogenous leukaemia and isolation of a fusion transcript, AML1/ETO, with similarity to Drosophila segmentation gene runt. Blood 80: 1825–1831PubMedGoogle Scholar
  20. Fan C-M, Kuwana E, Bulfone A, Fletcher CF, Copeland NG, Jenkins NA, Crews S, Martinez S, Puelles L, Rubenstein JLR, Tessier-Lavigne M (1996) 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 7: 1–16PubMedCrossRefGoogle Scholar
  21. Freidl M, Gulesserian T, Lubec G, Fountoulakis M, Lubec B (2001) Deterioration of the transcriptional, splicing and elongation machinery in brain of fetal Down syndrome. J Neural Transm [Suppl] 61: 47–57Google Scholar
  22. Gulesserian T, Engidawork E, Fountoulakis M, Lubec G (2001) Antioxidant proteins in fetal brain: superoxide dismutase-1 (SOD1) protein is not overexpressed in fetal Down syndrome. J Neural Transm [Suppl] 61: 71–84Google Scholar
  23. He X, Rosenfeld MG (1991) Mechanisms of complex transcriptional regulation: implications for brain development. Neuron 7: 183–196PubMedCrossRefGoogle Scholar
  24. Hewett PW, Nishi K, Daft EL, Clifford Murray J (2001) Selective expression of erg isoforms in human endothelial cells. IJBCB 33: 347–355Google Scholar
  25. Ianello RC, Crack PJ, de Haan JB, Kola I (1999) Oxidative stress and neural dysfunction in Down Syndrome. J Neural Transm [Suppl] 57: 257–267Google Scholar
  26. Ichikawa H, Shimizu K, Hayashi Y, Ohki M (1994) An RNA-binding protein gene, TLS/ FUS, is fused to ERG in human myeloid leukaemia with t(16;21) chromosomal translocation. Cancer Res 54: 2865–2868PubMedGoogle Scholar
  27. Ishii T, Itoh K, Takahashi S, Sato H, Yanagawa T, Katoh Y, Bannai S, Yamamoto M (2000) Transcription factor Nrf2 coordinately regulates a group of oxidative stress-inducible genes in macrophages. J Biol Chem 275: 16023–16029PubMedCrossRefGoogle Scholar
  28. Johnston MV, Alemi L, Harum KH (2003) Learning, memory, and transcription factors. Pediatr Res 53, 3: 369–374PubMedCrossRefGoogle Scholar
  29. Kanezaki R, Toki T, Yokoyama M, Yomogida K, Sugiyama K, Yamamoto M, Igarashi K, Ito E (2001) Transcription factor BACH1 is recruited to the nucleus by its novel alternative spliced isoform. J Biol Chem 276: 7278–7284PubMedCrossRefGoogle Scholar
  30. Keyse SM, Tyrrell RM (1989) Heme oxygenase is the major 32-kDa stress protein induced in human skin fibroblast by UVA radiation, hydrogen peroxide, and sodium arsenite. Proc Natl Acad Sci USA 86: 99–103PubMedCrossRefGoogle Scholar
  31. Kitamuro T, Takahashi K, Ogawa K, Udono RF, Takeda K, Furuyama K, Nakayama M, Sun J, Fujita H, Hida W, Hattori T, Shirato K, Igarashi K, Shibahara S (2003) Bachl functions as a hypoxia-inducible repressor for heme oxygenase-1 gene in human cells. J Biol Chem 278: 9125–9133PubMedCrossRefGoogle Scholar
  32. Labudova O, Krapfenbauer K, Moenkemann H, Rink H, Kitzmuller E, Cairns N, Lubec G (1998) Decreased transcription factor junD in brains of patients with Down syndrome. Neurosci Lett 252: 159–62PubMedCrossRefGoogle Scholar
  33. Labudova O, Kitzmueller E, Rink H, Cairns N, Lubec G (1999) Gene expression in fetal Down syndrome brain as revealed by substractive hybridization. J Neural Transm [Suppl] 59: 125–136Google Scholar
  34. Maroulakou IG, Bowe DB (2000) Expression and function of Ets transcription factors in mammalian development: a regulatory network. Oncogene 19: 6432–6442PubMedCrossRefGoogle Scholar
  35. Miyoshi H, Shimizu K, Kozu T, Maseki N, Kaneko Y, Ohki M (1991) t(8;21) breakpoints on chromosome 21 in acute myeloid leukaemia are clustered within a limited region of a single gene AML1. Proc Natl Acad Sci USA 88: 10431–10434CrossRefGoogle Scholar
  36. Morse D, Choi AMK (2002) Heme oxygenase-1, the “emerging molecule” has arrived. J Respir Cell Mol Biol 27: 8–16Google Scholar
  37. Ohira M, Seki N, Nagase T, Ishikawa K, Nomura N, Ohara O (1998) Characterization of a human homolog (BACH1) of the mouse Bachi gene encoding a BTB-Basic leucine zipper transcription factor and its mapping to chromosome 21q22.1. Genomics 47: 300–306PubMedCrossRefGoogle Scholar
  38. Otto F, Lübbert M, Stock M (2003) Upstream and downstream targets of RUNX proteins. J Cell Biochem 89: 9–18PubMedCrossRefGoogle Scholar
  39. Oyake T, Itoh K, Motohashi N, Hayashi N, Hoshino H, Nishizawa M, Yamamoto M, Igarashi K (1996) Bach proteins belong to a novel family of BTB-Basic leucine zipper transcription factors that interact with MafK and regulate transcription through the NF-E2 site. Mol Cell Biol 16: 6083–6095PubMedGoogle Scholar
  40. Perry Ch, Sklan EH, Birikh K, Shapira M, Trejo L, Eldor A, Soreq H (2002) Complex regulation of acetylcholinesterase gene expression in human brain tumors. Oncogene 21: 8428–8441PubMedCrossRefGoogle Scholar
  41. Poss KD, Tonegawa S (1997) Reduced stress defense in heme oxygenase 1-decficient cells. Proc Natl Acad Sci USA 94: 10925–10930PubMedCrossRefGoogle Scholar
  42. Reddy ES, Rao VN, Papas TS (1987) The erg gene: a human gene related to the ets oncogene. Proc Natl Acad Sci USA 84: 6131–6135PubMedCrossRefGoogle Scholar
  43. Su AI, Cooke MP, Ching KA, Hakak Y, Walker JR, Wiltshire T, Orth AP, Vega RG, Sapinoso LM, Morqrich A, Patapoutian A, Hampton GM, Schultz PG, Hogenesch JB (2002) Large-scale analysis of the human and mouse transcriptomes. Proc Natl Acad Sci USA 99: 4465–70PubMedCrossRefGoogle Scholar
  44. Sun J, Hoshino H, Takaku K, Nakajima O, Muto A, Suzuki H, Tashiro S, Takahashi S, Shibahara S, Alam J, Taketo MM, Yamamoto M, Igarashi K (2002) Hemoprotein Bachl regulates enhancer availability of heme oxygenase-1 gene. EMBO 21: 5216–5224CrossRefGoogle Scholar
  45. Tsuji K, Noda M (2000) Identification and expression of a novel 3’-exon of mouse Runx1/ Pebp2aB/Cbfa2/AML1 gene. Biochem Biophys Res Corn 274: 171–176CrossRefGoogle Scholar
  46. Vialard F, Toyama K, Vernoux S, Carlson EJ, Epstein CJ, Sinet PM, Rahmani Z (2000) Overexpression of mSim2 gene in the zona limitans of the diencephalons of segmental trisomy 16 Ts1Cje fetuses, a mouse model for trisomy 21: a novel whole-mount based RNA hybridisation study. Dev Brain Res 121: 73–78CrossRefGoogle Scholar
  47. Vlaeminck-Guillem V, Carrere S, Dewitte F, Stehelin D, Desbians X, DuterqueCoquillaud M (2000) The ets family member erg gene is expressed in mesodermal tissues and neural crests at fundamental steps during mouse embryogenesis. Mech Dev 91: 331–335PubMedCrossRefGoogle Scholar
  48. Weitzdoerfer R, Fountoulakis M, Lubec G (2002) Reduction of actin-related protein complex 2/3 in fetal Down Syndrome brain. Biochem Biophys Res Commun 293: 836–41PubMedCrossRefGoogle Scholar
  49. Yamaki A, Tochigi J, Kudoh J, Minoshima S, Shimizu N, Shimizu Y (2001) Molecular mechanisms of human single-minded 2 (SIM2) gene expression: identification of a promoter site in the SIM2 genomic sequence. Gene 270: 265–275PubMedCrossRefGoogle Scholar
  50. Yamashiro t, Aberg T, Levanon D, Groner Y, Thesleff I (2002) Expression of Runx-1, -2 and -3 during tooth, palate and craniofacial bone development. GEP 2: 109–112PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • R. Ferrando-Miguel
    • 1
  • M. S. Cheon
    • 1
  • J.-W. Yang
    • 1
  • G. Lubec
    • 1
    • 2
  1. 1.Department of PediatricsUniversity of ViennaAustria
  2. 2.CChem, FRSC (UK), Department of PediatricsUniversity of ViennaViennaAustria

Personalised recommendations