Reduction of nucleoside diphosphate kinase B, Rab GDP- dissociation inhibitor beta and histidine triad nucleotide-binding protein in fetal Down Syndrome brain

  • R. Weitzdoerfer
  • D. Stolzlechner
  • M. Dierssen
  • J. Ferreres
  • M. Fountoulakis
  • G. Lubec


Information on the various factors leading to impairments in the developing brain of fetal Down Syndrome patients is limited to few histological reports. We therefore attempted to describe expression levels of proteins in brain using the proteomic technique of two-dimensional electrophoresis with subsequent mass spectroscopical identification of protein spots and quantification with specific software. Cortical tissue was obtained from autopsy of human fetal abortus. Protein levels of GTP-binding nuclear protein ran, guanine nucleotide-binding protein g(o), alpha subunit 2, guanine nucleotide-binding protein g(i)/g(s)/g(t) beta subunit 1, -beta subunit 2, guanine nucleotide-binding protein beta subunit 5, nucleoside diphosphate kinase A, nucleoside diphosphate kinase B, Rab GDP-dissociation inhibitor beta, Rho GDP-dissociation inhibitor 1, biphosphate 3′-nucleotidase, small glutamine-rich tetra-tricopeptide repeat-containing protein and histidine triad nucleotide-binding protein were studied.

Quantification revealed statistically significant reduced levels of nucleoside diphosphate kinase B, Rab GDP-dissociation inhibitor beta and histidine triad nucleotide-binding protein in fetal DS brain as compared to controls.

We conclude that in early prenatal life proteins involved in neural differentiation, migration and synaptic transmission are impaired in DS cortex. These results may help to understand the abundant mechanisms leading to abnormalities in the wiring, structure and function of DS brain.


Down Syndrome Nucleoside Diphosphate Kinase Down Syndrome Patient Neuropathy Target Esterase Nm23 Gene 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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  1. Agou F, Raveh S, Mesnildrey S, Veron S (1999) Single strand DNA specificity analysis of human nucleoside diphosphate kinase B. J Biol Chem 274: 19630–19636PubMedCrossRefGoogle Scholar
  2. Backer J, Mendola C, Kovesdi I, Fairhurst J, O’Hara B, Eddy RL, Shows T, Mathew S, Murty V, Chaganti R (1993) Chromosomal localisation and nucleoside diphosphate kinase activity of human metastasis-suppressor genes NM23-1 and NM23-2. Oncogene 8: 497–502PubMedGoogle Scholar
  3. Berndt P, Hobolm U, Langen H (1999) Reliable automatic protein identification from matrix-assisted laser desorption/ionisation mass spectrometric peptide fingerprints.Electrophoresis 20: 3521–3526PubMedCrossRefGoogle Scholar
  4. Bradford M (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
  5. Brzoska P, Chen H, Levin N, Kuo WL, Collins C, Fu KK, Gray J, Christman M (1996) Cloning, mapping and in vivo localisation of a human member of the PKCI-1 protein family (PKCNH1). Genomics 36: 151–156PubMedCrossRefGoogle Scholar
  6. D’Adamo P, Menegon A, Lo Nigro C, Grasso M, Tamanini F, Bienvenu T, Gedeon A,Oostra B, Wu SK, Tandon A, Valtorta F, Balch W, Chelly J, Toniolo D (1998) Mutations in GDI1 are responsible for X-linked non-specific mental retardation.Nature Genet 19: 134–139PubMedCrossRefGoogle Scholar
  7. De la Monte S (1999) Molecular abnormalities of the brain in Down syndrome:relevance to Alzheimer’s neurodegeneration. J Neural Transm [Suppl] 57: 1–20Google Scholar
  8. Dierssen M, Vallina I, Baamonde C, Garcia-Calatayud S, Lumbreras M, Florez J (1997) Alterations of central noradrenergic transmission in Ts65Dn mouse, a model for Down syndrome. Brain Res 749: 238–244PubMedCrossRefGoogle Scholar
  9. Epstein C (1995) The metabolic and molecular bases of inherited disease In: Scriver SR,Beaudet AL, Sly WS, Valle D (eds) Down Syndrome (Trisomy 21). McGraw Hill,New York, pp 749–794Google Scholar
  10. Fleig U, Salus S, Karig I, Sazer S (2000) The fission yeast ran GTPase is required for microtubule integrity. J Cell Biol 151: 1101–1112PubMedCrossRefGoogle Scholar
  11. Fountoulakis M, Langen H (1997) Identification of proteins by matrix-assisted laser desorption ionisation-mass spectrometry following in-gel digestion in low-salt, non- volatile buffer and simplified peptide recovery. Anal Biochem 250: 153–156PubMedCrossRefGoogle Scholar
  12. Fountoulakis M, Cairns N, Lubec G (1999) Increased levels of 14-3-3 gamma and epsilon proteins in brain of patients with Alzheimer’s disease and Down Syndrome. J Neural Transm [Suppl] 57: 323–335Google Scholar
  13. Fuentes J, Genesca L, Kingsbury T, Cunningham K, Perez-Riba M, Estivill X, de la Luna S (2000) DSCR1, over expressed in Down Syndrome, is an inhibitor of calcineurin-mediated signalling pathways. Hum Mol Genet 9: 1681–1690PubMedCrossRefGoogle Scholar
  14. Gilman A (1987) G proteins: transducers of receptor-generated signals. Annu Rev Biochem 56: 615–649PubMedCrossRefGoogle Scholar
  15. Glynn P (2000) Neural development and neurodegeneration: two faces of neuropathy target esterase. Prog Neurobiol 61: 61–74PubMedCrossRefGoogle Scholar
  16. Joosten M, Vankan-Berkhoudt Y, Van den Broek M, Beijen A, Lowenberg B, von Lindern M, Deiwel R (2000) The gene NM23-M2 is frequently mutated in Leukemia.Exp Hematol 28: 1491CrossRefGoogle Scholar
  17. Kaziro Y (1978) The role of guanosine 5’-triphosphate in polypeptide chain elongation.Biochim Biophys Acta 505: 95–127PubMedCrossRefGoogle Scholar
  18. Keim D, Hailat N, Melhem R, Zhu XX, Lascu I, Veron M, Strahler J, Hanasch S (1992) Proliferation-related expression of pl9/nm23 nucleoside diphosphate kinase. J Clin Invest 89: 919–924PubMedCrossRefGoogle Scholar
  19. Korsisaari N, Mäkelä T (2000) Interaction of Cdk7 and Kin28 with Hint/PKCI-1 and Hntl Histidine triad proteins. J Biol Chem 275: 34837–34840PubMedCrossRefGoogle Scholar
  20. Kraeft S, Traincart F, Bourdais J, Mesnildrey S, Veron M, Chen LB (1996) Nuclear localisation of nucleoside diphosphate kinase type B (nm23-H2) in cultured cells. Exp Cell Res 246: 355–367Google Scholar
  21. Langen H, Roder D, Juranville J, Fountoulakis M (1997) Effect of protein application mode and acrylamide concentration on the resolution of protein spots separated by two-dimensional gel electrophoresis. Electrophoresis 18: 2085–2090PubMedCrossRefGoogle Scholar
  22. Lima C, Klein M, Hendrickson W (1997) Structure-based analysis of catalysis and substrate definition in the HIT protein family. Science 278: 286–290PubMedCrossRefGoogle Scholar
  23. Nuoffer C, Davidson HW, Matteson J, Meinkoth J, Balch W (1994) A GDP-bound of rabl inhibits protein export from the endoplasmic reticulum and transport between Golgi compartments. J Cell Biol 125: 225–237PubMedCrossRefGoogle Scholar
  24. Ohneda K, Fukuda M, Shimada N, Ishikawa N, Ichou T, Kaji K, Toyota T, Kimura N (1994) Increased expression of nucleoside diphosphate kinases/nm23 in human diploid fibroblasts transformed by SV40 large T antigen or 60Co irradiation. FEBS Lett 348: 273–277PubMedCrossRefGoogle Scholar
  25. Okabe-Kado J, Kasukabe T, Hozumi M, Honma Y, Kimura N, Baba H, Urano T, Shiku H (1995) A new function of Nm23/NDP kinase as a differentiation inhibitory factor, which does not require its kinase activity. FEBS Lett 363: 311–315PubMedCrossRefGoogle Scholar
  26. Postel E (1999) Cleavage of DNA by human NM23-H2/Nucleoside diphosphate kinase involves formation of a covalent protein-DNA complex. J Biol Chem 274: 22821–22829PubMedCrossRefGoogle Scholar
  27. Postel E, Berberich S, Flint S, Ferrone C (1993) Human c-myc transcription factor PuF identified as nm23-H2 nucleoside diphosphate kinase, a candidate suppressor of tumor metastasis. Science 261: 478–480PubMedCrossRefGoogle Scholar
  28. Postel E, Weiss V, Beneken J, Kirtane A (1996) Mutational analysis of NM23-H2/NDP kinase identifies the structural domains critical to recognition of a c-myc regulatory element. Proc Natl Acad Sci USA 93: 6892–6897PubMedCrossRefGoogle Scholar
  29. Postel E, Abramczyk B, Levit M, Kyin S (2000) Catalysis of DNA cleavage and nucleoside triphosphate synthesis by NM″§-H2/NDP kinase share an active site that implies a DNA repair function. Proc Natl Acad Sci USA 97: 14194–14199PubMedCrossRefGoogle Scholar
  30. Raveh S, Vinh J, Rossier J, Agou F, Veron M (2001) Peptidic determinants and structural model of human NDP kinase B (Nm23-H2) bound to single-stranded DNA. Biochem 40: 5882–5893CrossRefGoogle Scholar
  31. Razin E, Zhang ZC, Nechushtan H, Frenkel S, Lee YN, Arudchandran R, Rivera J (1999) Suppression of Microphthalmia transcriptional activity by its association with Protein kinase C-interacting Protein 1 in mastcells. J Biol Chem 274: 34272–34276PubMedCrossRefGoogle Scholar
  32. Reed W, Ozand P (1985) Adrenergic sensitivity of cultured murine trisomy 16 cells. Dev Pharmacol Ther 8: 200–208PubMedGoogle Scholar
  33. Roymans D, Willems R, Vissenberg K, De Jonghe C, Grobben B, Claes P, Lascu I, Van Bockstaele D, Verbelen J, Van Broeckhoven C, Siegers H (2000) Nucleoside diphosphate kinase beta (Nm23-Rl/NDPKbeta) is associated with intermediate filaments and becomes upregulated upon cAMP-induced differentiation of rat C6 glioma. Exp Cell Res 261: 127–138PubMedCrossRefGoogle Scholar
  34. Sanes JR, Jessell TM (2000) The guidance of axons to their targets. In: Kandel ER, Schwartz JH, Jessell TM (eds) Principles of neural science. McGraw Hill, New York, pp 1063–1086Google Scholar
  35. Schimmoller F, Simon I, Pfeffer S (1998) Rab GTPases, Directors of vesicle docking. J Biol Chem 273: 22161–22164PubMedCrossRefGoogle Scholar
  36. Schubert P, Morino T, Miyazaki H, Ogata T, Nakamura Y, Marchini C, Ferroni S (2000) Cascading glia reactions: a common pathomechanism and its differentiated control by cyclic nucleotide signalling. Ann NY Acad Sci 903: 24–33PubMedCrossRefGoogle Scholar
  37. Siprashvili Z, Sozzi G, Barnes L, Mc Cue P, Robinson A, Eryomin V, Sard L, Tagliabue E, Greco A, Fusetti L, Schwartz G, Pierotti M, Croce C, Huebner K (1997) Replacement of FHIT in cancer cells suppresses tumorigenicity. Proc Natl Acad Sci USA 94: 13771–13776PubMedCrossRefGoogle Scholar
  38. Shisheva A, Chinni S, DeMarco C (1999) general role of GDP Dissociation Inhibitor 2 in membrane release of Rab proteins: modulation of its functional interactions by in vitro and in vivo structural modifications. Biochem 38: 11711–11721CrossRefGoogle Scholar
  39. Spiegelberg B, Xiong JP, Smith JJ, Gu R, York J (1999) Cloning and characterization of a mammalian lithium-sensitive bisphosphate 3’-nucleotidase inhibited by inositol 1,4-bisphosphate. J Biol Chem 274: 13619–13628PubMedCrossRefGoogle Scholar
  40. Stahl J, Leone A, Rosengard A, Porter L, King C, Steeg P (1991) Identification of a second human nm23 gene, nm23-H2. Cancer Res 51: 445–449PubMedGoogle Scholar
  41. Takai Y, Sasaki T, Matozaki T (2001) Small GTP-binding proteins. Physiol Rev 81: 153–208PubMedGoogle Scholar
  42. Venturelli D, Martinez R, Melotti P, Casella I, Peschle C, Cucco C, Spampinato G, Darzynkiewicz Z, Calabretta B (1995) Overexpression of DR-nm23, a protein encoded by a member of the nm23 gene family, inhibits granulocyte differentiation and induces apoptosis in 32Dcl3 myeloid cells. Proc Natl Acad Sci USA 92: 7435–7439PubMedCrossRefGoogle Scholar
  43. Wisniewski K, Kida E (1994) Abnormal neurogenesis and synaptogenesis in Down Syndrome brain. Dev Brain Dysfunct 7: 289–301Google Scholar

Copyright information

© Springer-Verlag/Wien 2001

Authors and Affiliations

  • R. Weitzdoerfer
    • 1
  • D. Stolzlechner
    • 1
  • M. Dierssen
    • 2
  • J. Ferreres
    • 2
  • M. Fountoulakis
    • 3
  • G. Lubec
    • 1
  1. 1.Department of PediatricsUniversity of ViennaViennaAustria
  2. 2.Medical and Molecular Genetics Center-IROHospital Duran i ReynalsBarcelonaSpain
  3. 3.Gene TechnologiesF. Hoffmann-La RocheBaselSwitzerland

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