Molecular Genetics and Genomics

, Volume 283, Issue 2, pp 147–156 | Cite as

Null mutations in Drosophila Optomotor-blind affect T-domain residues conserved in all Tbx proteins

  • Aditya Sen
  • Christian Gadomski
  • Jürgen Balles
  • Yasmin Abassi
  • Christian Dorner
  • Gert O. PflugfelderEmail author
Original Paper


The T-box transcription factors TBX2 and TBX3 are overexpressed in many human cancers raising the need for a thorough understanding of the cellular function of these proteins. In Drosophila, there is one corresponding ortholog, Optomotor-blind (Omb). Currently, only two missense mutations are known for the two human proteins. Making use of the developmental defects caused by inactivation of omb, we have isolated and molecularly characterized four new omb mutations, three of them are missense mutations of amino acids fully conserved in all Tbx proteins. We interpret the functional defects in the framework of the known structure of the human TBX3 protein and provide evidence for loss of Omb DNA-binding activity in all three newly identified missense mutations.


T-box (Tbx) optomotor-blind Mutation Missense Protein structure DNA binding Drosophila 



We thank the Bloomington Stock Center for Drosophila stocks, E. Jost for fly care, and Deutsche Forschungsgemeinschaft for grant Pf163/14-1.

Supplementary material

438_2009_505_MOESM1_ESM.doc (48 kb)
Supplementary material 1 (DOC 47 kb)


  1. Andreou AM, Pauws E, Jones MC, Singh MK, Bussen M, Doudney K, Moore GE, Kispert A, Brosens JJ, Stanier P (2007) TBX22 missense mutations found in patients with X-linked cleft palate affect DNA binding, sumoylation, and transcriptional repression. Am J Hum Genet 81:700–712CrossRefPubMedGoogle Scholar
  2. Ashburner M (1989) Drosophila. A laboratory handbook. Cold Spring Harbor Laboratory Press, Cold Spring HarborGoogle Scholar
  3. Bamshad M, Lin RC, Law DJ, Watkins WS, Krakowiak PA, Moore ME, Fransceschini P, Lala R, Holmes LB, Gebuhr TC, Bruneau BG, Schinzel A, Seidman JG, Seidman CE, Jorde LB (1997) Mutations in human TBX3 alter limb, apocrine and genital development in ulnar-mammary syndrome. Nat Genet 16:311–315CrossRefPubMedGoogle Scholar
  4. Bamshad M, Le T, Watkins WS, Dixon ME, Kramer BE, Roeder AD, Carey JC, Root S, Schinzel A, Van Maldergem L, Gardner RJ, Lin RC, Seidman CE, Seidman JG, Wallerstein R, Moran E, Sutphen R, Campbell CE, Jorde LB (1999) The spectrum of mutations in TBX3: genotype/phenotype relationship in ulnar-mammary syndrome. Am J Hum Genet 64:1550–1562CrossRefPubMedGoogle Scholar
  5. Basson CT, Bachinsky DR, Lin RC, Levi T, Elkins JA, Soults J, Grayzel D, Kroumpouzou E, Traill TA, Leblanc-Straceski J, Renault B, Kucherlapati R, Seidman JG, Seidman CE (1997) Mutations in human cause limb and cardiac malformation in Holt-Oram syndrome. Nat Genet 15:30–34CrossRefPubMedGoogle Scholar
  6. Basson CT, Huang T, Lin RC, Bachinsky DR, Weremowicz S, Vaglio A, Bruzzone R, Quadrelli R, Lerone M, Romeo G, Silengo M, Pereira A, Krieger J, Mesquita SF, Kamisago M, Morton CC, Pierpont ME, Muller CW, Seidman JG, Seidman CE (1999) Different TBX5 interactions in heart and limb defined by Holt-Oram syndrome mutations. Proc Natl Acad Sci USA 96:2919–2924CrossRefPubMedGoogle Scholar
  7. Bilican B, Goding CR (2006) Cell cycle regulation of the T-box transcription factor tbx2. Exp Cell Res 312:2358–2366CrossRefPubMedGoogle Scholar
  8. Bollag RJ, Siegfried Z, Cebra-Thomas JA, Garvey N, Davison EM, Silver LM (1994) An ancient family of embryonically expressed mouse genes sharing a conserved protein motif with the T locus. Nat Genet 7:383–389CrossRefPubMedGoogle Scholar
  9. Bongers EM, Duijf PH, van Beersum SE, Schoots J, Van Kampen A, Burckhardt A, Hamel BC, Losan F, Hoefsloot LH, Yntema HG, Knoers NV, van Bokhoven H (2004) Mutations in the human TBX4 gene cause small patella syndrome. Am J Hum Genet 74:1239–1248CrossRefPubMedGoogle Scholar
  10. Brassington AM, Sung SS, Toydemir RM, Le T, Roeder AD, Rutherford AE, Whitby FG, Jorde LB, Bamshad MJ (2003) Expressivity of Holt-Oram syndrome is not predicted by TBX5 genotype. Am J Hum Genet 73:74–85CrossRefPubMedGoogle Scholar
  11. Braybrook C, Doudney K, Marcano AC, Arnason A, Bjornsson A, Patton MA, Goodfellow PJ, Moore GE, Stanier P (2001) The T-box transcription factor gene TBX22 is mutated in X-linked cleft palate and ankyloglossia. Nat Genet 29:179–183CrossRefPubMedGoogle Scholar
  12. Brunner A, Wolf R, Pflugfelder GO, Poeck B, Heisenberg M (1992) Mutations in the proximal region of the optomotor-blind locus of Drosophila melanogaster reveal a gradient of neuroanatomical and behavioural phenotypes. J. Neurogenet 8:43–55CrossRefPubMedGoogle Scholar
  13. Brunner E, Brunner D, Fu W, Hafen E, Basler K (1999) The dominant mutation Glazed is a gain-of-function allele of wingless that, similar to loss of APC, interferes with normal eye development. Dev Biol 206:178–188CrossRefPubMedGoogle Scholar
  14. Carlson H, Ota S, Campbell CE, Hurlin PJ (2001) A dominant repression domain in Tbx3 mediates transcriptional repression and cell immortalization: relevance to mutations in Tbx3 that cause ulnar-mammary syndrome. Hum Mol Genet 10:2403–2413CrossRefPubMedGoogle Scholar
  15. Cartegni L, Chew SL, Krainer AR (2002) Listening to silence and understanding nonsense: exonic mutations that affect splicing. Nat Rev Genet 3:285–298CrossRefPubMedGoogle Scholar
  16. Chaabouni M, Smaoui N, Benneji N, M’Rad R, Jemaa LB, Hachicha S, Chaabouni H (2005) Mutation analysis of TBX22 reveals new mutation in Tunisian CPX family. Clin Dysmorphol 14:23–25CrossRefPubMedGoogle Scholar
  17. Coll M, Seidman JG, Müller CW (2002) Structure of the DNA-bound T-box domain of human TBX3, a transcription factor responsible for ulnar-mammary syndrome. Structure (Camb) 10:343–356CrossRefGoogle Scholar
  18. Fan C, Liu M, Wang Q (2003) Functional analysis of TBX5 missense mutations associated with Holt-Oram syndrome. J Biol Chem 278:8780–8785CrossRefPubMedGoogle Scholar
  19. Garnett AT, Han TM, Gilchrist MJ, Smith JC, Eisen MB, Wardle FC, Amacher SL (2009) Identification of direct T-box target genes in the developing zebrafish mesoderm. Development 136:749–760CrossRefPubMedGoogle Scholar
  20. Ghosh TK, Packham EA, Bonser AJ, Robinson TE, Cross SJ, Brook JD (2001) Characterization of the TBX5 binding site and analysis of mutations that cause Holt-Oram syndrome. Hum Mol Genet 10:1983–1994CrossRefPubMedGoogle Scholar
  21. Gray PA, Fu H, Luo P, Zhao Q, Yu J, Ferrari A, Tenzen T, Yuk DI, Tsung EF, Cai Z, Alberta JA, Cheng LP, Liu Y, Stenman JM, Valerius MT, Billings N, Kim HA, Greenberg ME, McMahon AP, Rowitch DH, Stiles CD, Ma Q (2004) Mouse brain organization revealed through direct genome-scale TF expression analysis. Science 306:2255–2257CrossRefPubMedGoogle Scholar
  22. Grimm S, Pflugfelder GO (1996) Control of the gene optomotor-blind in Drosophila wing development by decapentaplegic and wingless. Science 271:1601–1603CrossRefPubMedGoogle Scholar
  23. Habets PE, Moorman AF, Clout DE, van Roon MA, Lingbeek M, van Lohuizen M, Campione M, Christoffels VM (2002) Cooperative action of Tbx2 and Nkx2.5 inhibits ANF expression in the atrioventricular canal: implications for cardiac chamber formation. Genes Dev 16:1234–1246CrossRefPubMedGoogle Scholar
  24. Halder G, Carroll SB (2001) Binding of the Vestigial co-factor switches the DNA-target selectivity of the Scalloped selector protein. Development 128:3295–3305PubMedGoogle Scholar
  25. Heinritz W, Moschik A, Kujat A, Spranger S, Heilbronner H, Demuth S, Bier A, Tihanyi M, Mundlos S, Gruenauer-Kloevekorn C, Froster UG (2005a) Identification of new mutations in the TBX5 gene in patients with Holt-Oram syndrome. Heart 91:383–384CrossRefPubMedGoogle Scholar
  26. Heinritz W, Shou L, Moschik A, Froster UG (2005b) The human TBX5 gene mutation database. Hum Mutat 26:397CrossRefPubMedGoogle Scholar
  27. Hiroi Y, Kudoh S, Monzen K, Ikeda Y, Yazaki Y, Nagai R, Komuro I (2001) Tbx5 associates with Nkx2-5 and synergistically promotes cardiomyocyte differentiation. Nat Genet 28:276–280CrossRefPubMedGoogle Scholar
  28. Hofmeyer K, Kretzschmar D, Pflugfelder G (2008) optomotor-blind expression in glial cells is required for correct axonal projection across the Drosophila inner optic chiasm. Dev Biol 315:28–41CrossRefPubMedGoogle Scholar
  29. Ivanova N, Dobrin R, Lu R, Kotenko I, Levorse J, DeCoste C, Schafer X, Lun Y, Lemischka IR (2006) Dissecting self-renewal in stem cells with RNA interference. Nature 442:533–538CrossRefPubMedGoogle Scholar
  30. Kerscher S, Albert S, Wucherpfennig D, Heisenberg M, Schneuwly S (1995) Molecular and genetic analysis of the Drosophila mas-1 (mannosidase-1) gene which encodes a glycoprotein processing a-1,2 mannosidase. Dev Biol 168:613–626CrossRefPubMedGoogle Scholar
  31. Kirk EP, Sunde M, Costa MW, Rankin SA, Wolstein O, Castro ML, Butler TL, Hyun C, Guo G, Otway R, Mackay JP, Waddell LB, Cole AD, Hayward C, Keogh A, Macdonald P, Griffiths L, Fatkin D, Sholler GF, Zorn AM, Feneley MP, Winlaw DS, Harvey RP (2007) Mutations in cardiac T-box factor gene TBX20 are associated with diverse cardiac pathologies, including defects of septation and valvulogenesis and cardiomyopathy. Am J Hum Genet 81:280–291CrossRefPubMedGoogle Scholar
  32. Kispert A, Herrmann BG (1993) The Brachyury gene encodes a novel DNA binding protein. EMBO J 12:3211–3220PubMedGoogle Scholar
  33. Kispert A, Koschorz B, Herrmann BG (1995) The T protein encoded by Brachyury is a tissue-specific transcription factor. EMBO J 14:4763–4772PubMedGoogle Scholar
  34. Kopp A, Duncan I (1997) Control of cell fate and polarity in the abdominal segments of Drosophila by optomotor-blind. Development 124:3715–3726PubMedGoogle Scholar
  35. Krause A, Zacharias W, Camarata T, Linkhart B, Law E, Lischke A, Miljan E, Simon HG (2004) Tbx5 and Tbx4 transcription factors interact with a new chicken PDZ-LIM protein in limb and heart development. Dev Biol 273:106–120CrossRefPubMedGoogle Scholar
  36. Lausch E, Hermanns P, Farin HF, Alanay Y, Unger S, Nikkel S, Steinwender C, Scherer G, Spranger J, Zabel B, Kispert A, Superti-Furga A (2008) TBX15 mutations cause craniofacial dysmorphism, hypoplasia of scapula and pelvis, and short stature in Cousin syndrome. Am J Hum Genet 83:649–655CrossRefPubMedGoogle Scholar
  37. Lingbeek ME, Jacobs JJ, van Lohuizen M (2002) The T-box repressors TBX2 and TBX3 specifically regulate the tumor suppressor gene p14ARF via a variant T-site in the initiator. J Biol Chem 277:26120–26127CrossRefPubMedGoogle Scholar
  38. Liu C, Shen A, Li X, Jiao W, Zhang X, Li Z (2008) T-box transcription factor TBX20 mutations in Chinese patients with congenital heart disease. Eur J Med Genet 51:580–587CrossRefPubMedGoogle Scholar
  39. Macindoe I, Glockner L, Vukasin P, Stennard FA, Costa MW, Harvey RP, Mackay JP, Sunde M (2009) Conformational stability and DNA binding specificity of the cardiac T-box transcription factor Tbx20. J Mol Biol 389:606–618Google Scholar
  40. Marcano AC, Doudney K, Braybrook C, Squires R, Patton MA, Lees MM, Richieri-Costa A, Lidral AC, Murray JC, Moore GE, Stanier P (2004) TBX22 mutations are a frequent cause of cleft palate. J Med Genet 41:68–74CrossRefPubMedGoogle Scholar
  41. Meneghini V, Odent S, Platonova N, Egeo A, Merlo GR (2006) Novel TBX3 mutation data in families with Ulnar-Mammary syndrome indicate a genotype-phenotype relationship: mutations that do not disrupt the T-domain are associated with less severe limb defects. Eur J Med Genet 49:151–158CrossRefPubMedGoogle Scholar
  42. Minguillon C, Logan M (2003) The comparative genomics of T-box genes. Brief Funct Genom Proteom 2:224–233CrossRefGoogle Scholar
  43. Miyahara K, Suzuki N, Ishihara T, Tsuchiya E, Katsura I (2004) TBX2/TBX3 transcriptional factor homologue controls olfactory adaptation in Caenorhabditis elegans. J Neurobiol 58:392–402CrossRefPubMedGoogle Scholar
  44. Morgan TH, Bridges CB (1916) Sex-linked inheritance in Drosophila. Carnegie Institution of Washington Publication no. 237, pp 1–88Google Scholar
  45. Müller CW, Herrmann BG (1997) Crystallographic structure of the T domain-DNA complex of the Brachyury transcription factor. Nature 389:884–888CrossRefPubMedGoogle Scholar
  46. Ng PC, Henikoff S (2006) Predicting the effects of amino acid substitutions on protein function. Annu Rev Genom Hum Genet 7:61–80CrossRefGoogle Scholar
  47. Packham EA, Brook JD (2003) T-box genes in human disorders. Hum Mol Genet 12:R37–R44CrossRefPubMedGoogle Scholar
  48. Papaioannou VE (2001) T-box genes in development: from hydra to humans. Int Rev Cytol 207:1–70CrossRefPubMedGoogle Scholar
  49. Pflugfelder GO (2009) omb and circumstance. J Neurogenet 23:15–33CrossRefPubMedGoogle Scholar
  50. Pflugfelder GO, Schwarz H, Roth H, Poeck B, Sigl A, Kerscher S, Jonschker B, Pak WL, Heisenberg M (1990) Genetic and molecular characterization of the optomotor-blind gene locus in Drosophila melanogaster. Genetics 126:91–104PubMedGoogle Scholar
  51. Pflugfelder GO, Roth H, Poeck B (1992a) A homology domain shared between Drosophila melanogaster and mouse Brachyury is involved in DNA binding. Biochem Biophys Res Commun 186:918–925CrossRefPubMedGoogle Scholar
  52. Pflugfelder GO, Roth H, Poeck B, Kerscher S, Schwarz H, Jonschker B, Heisenberg M (1992b) The lethal(1)optomotor-blind gene of Drosophila melanogaster is a major organizer of optic lobe development: isolation and characterization of the gene. Proc Natl Acad Sci USA 89:1199–1203CrossRefPubMedGoogle Scholar
  53. Poeck B, Balles J, Pflugfelder GO (1993) Transcript identification in the optomotor-blind locus of Drosophila melanogaster by intragenic recombination mapping and PCR-aided sequence analysis of lethal point mutations. Mol Gen Genet 238:325–332CrossRefPubMedGoogle Scholar
  54. Porsch M, Hofmeyer K, Bausenwein BS, Grimm S, Weber BHF, Miassod R, Pflugfelder GO (1998) Isolation of a Drosophila T-box gene closely related to human TBX1. Gene 212:237–248CrossRefPubMedGoogle Scholar
  55. Porsch M, Sauer M, Schulze S, Bahlo A, Roth M, Pflugfelder GO (2005) The relative role of the T-domain and flanking sequences for developmental control and transcriptional regulation in protein chimeras of Drosophila OMB and ORG-1. Mech Dev 122:81–96CrossRefPubMedGoogle Scholar
  56. Pulichino AM, Vallette-Kasic S, Couture C, Gauthier Y, Brue T, David M, Malpuech G, Deal C, Van Vliet G, De Vroede M, Riepe FG, Partsch CJ, Sippell WG, Berberoglu M, Atasay B, Drouin J (2003) Human and mouse TPIT gene mutations cause early onset pituitary ACTH deficiency. Genes Dev 17:711–716CrossRefPubMedGoogle Scholar
  57. Reamon-Buettner SM, Borlak J (2004) TBX5 mutations in non-Holt-Oram syndrome (HOS) malformed hearts. Hum Mutat 24:104CrossRefPubMedGoogle Scholar
  58. Rodriguez M, Aladowicz E, Lanfrancone L, Goding CR (2008) Tbx3 represses e-cadherin expression and enhances melanoma invasiveness. Cancer Res 68:7872–7881CrossRefPubMedGoogle Scholar
  59. Ryder E, Blows F, Ashburner M, Bautista-Llacer R, Coulson D, Drummond J, Webster J, Gubb D, Gunton N, Johnson G, O’Kane CJ, Huen D, Sharma P, Asztalos Z, Baisch H, Schulze J, Kube M, Kittlaus K, Reuter G, Maroy P, Szidonya J, Rasmuson-Lestander A, Ekstrom K, Dickson B, Hugentobler C, Stocker H, Hafen E, Lepesant JA, Pflugfelder G, Heisenberg M, Mechler B, Serras F, Corominas M, Schneuwly S, Preat T, Roote J, Russell S (2004) The DrosDel collection: a set of P-element insertions for generating custom chromosomal aberrations in Drosophila melanogaster. Genetics 167:797–813CrossRefPubMedGoogle Scholar
  60. Sasaki G, Ogata T, Ishii T, Hasegawa T, Sato S, Matsuo N (2002) Novel mutation of TBX3 in a Japanese family with ulnar-mammary syndrome: implication for impaired sex development. Am J Med Genet 110:365–369CrossRefPubMedGoogle Scholar
  61. Shen J, Dahmann C (2005) The role of Dpp signaling in maintaining the Drosophila anteroposterior compartment boundary. Dev Biol 279:31–43CrossRefPubMedGoogle Scholar
  62. Shen J, Dorner C, Bahlo A, Pflugfelder GO (2008) optomotor-blind suppresses instability at the A/P compartment boundary of the Drosophila wing. Mech Dev 125:233–246CrossRefPubMedGoogle Scholar
  63. Suphapeetiporn K, Tongkobpetch S, Siriwan P, Shotelersuk V (2007) TBX22 mutations are a frequent cause of non-syndromic cleft palate in the Thai population. Clin Genet 72:478–483PubMedCrossRefGoogle Scholar
  64. Tada M, Smith JC (2001) T-targets: clues to understanding the functions of T-box proteins. Dev Growth Differ 43:1–11CrossRefPubMedGoogle Scholar
  65. Tomlinson A (2003) Patterning the peripheral retina of the fly: decoding a gradient. Dev Cell 5:799–809CrossRefPubMedGoogle Scholar
  66. Vallette-Kasic S, Couture C, Balsalobre A, Gauthier Y, Metherell L, Dattani M, Drouin J (2007) The TPIT gene mutation M86R associated with isolated adrenocorticotropin deficiency interferes with protein: protein interactions. J Clin Endocrinol Metab 92:3991–3999CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Aditya Sen
    • 1
    • 3
  • Christian Gadomski
    • 1
  • Jürgen Balles
    • 2
  • Yasmin Abassi
    • 1
  • Christian Dorner
    • 1
  • Gert O. Pflugfelder
    • 1
    • 2
    Email author
  1. 1.Institute of GeneticsJohannes Gutenberg-UniversitätMainzGermany
  2. 2.Theodor Boveri-InstituteUniversität WürzburgWürzburgGermany
  3. 3.Department of Biochemistry and Molecular BiologyUniformed Services UniversityBethesdaUSA

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