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2006-12-18 - Colloque/Présentation - poster - Français - 1 page(s)

Tassin Alexandra , Ansseau Eugénie , Sauvage Sébastien, Mattéotti Christel, Van Acker Anne-Marie, Figlewicz Denise, Laoudj-Chenivesse Dalila, Oberdan Léo, Coppée Frédérique , Belayew Alexandra , "Evidence for the expression of the DUX4 gene in primary myoblasts of patients affected with Facioscapulohumeral muscular dystrophy" in 194th Congrès de la Société Belge de Biochimie et de Biologie Moléculaire / “Young Scientists Day , Gembloux, Belgique, 2006

  • Codes CREF : Biologie moléculaire (DI3111), Pathologies particulières (DI3370)
  • Unités de recherche UMONS : Biologie moléculaire (M122)
  • Instituts UMONS : Institut des Sciences et Technologies de la Santé (Santé)

Abstract(s) :

(Anglais) FSHD is an inherited dominant disease characterized by progressive weakness and atrophy of the muscles from the face to the lower limbs. The 4q35 locus linked to FSHD contains varying copy numbers of a 3.3-kb repeated element (D4Z4). This repeat array contracts from 11-100 copies in non- affected individuals down to 1-10 copies in patients, inducing a chromatin change that affects expression of several genes (VAN DER MAAREL et al, 2006). Our group has identified a double homeobox gene (DUX4) within each D4Z4 element (GABRIELS et al, 1999) but demonstration of its expression was technically challenging because of its low levels, toxicity, and homology to hundreds of DUX genes unlinked to FSHD. A similar gene (DUX4c) maps 42 kb centromeric of the repeat array (COPPEE et al, 2005) and expresses a protein we could detect by western blot in myoblasts and muscle biopsies, at higher levels in FSHD samples. The DUX4 and DUX4c proteins are 424- and 374- residue long, respectively, and are identical over their first 342-residues including the double homeodomain. The DUX4 carboxyl-terminal domain (“DUX4 tail”, 297 residues) was expressed as a His-tag fusion in E. coli, purified on Zn++-agarose beads (Affiland) and used to raise the 9A12 monoclonal antibody. It reacted specifically with DUX4 and DUX4c by immunoprecipitation of radioactive proteins expressed in vitro, western blotting performed on extracts of cells transfected with pCIneo-DUX expression vectors, immunofluorescence on transfected cells, and competition with antigen excess. Western blot was performed with 9A12 on extracts of human primary myoblast lines derived from muscle biopsie. Using a high sensitivity substrate (Femto, Pierce) we detected DUX4 in 8 FSHD myoblast lines but not in 3 controls. In addition, part of a DUX4 mRNA comprising the full open reading frame could be amplified by RT-PCR and confirmed by sequence determination in 4 FSHD lines but not in 2 controls. We acknowledge funding by the ABMM (Belgium), AFM (France), MDA and NIH (USA), and the Frenzel (Germany) and Shaw-Fischer (USA) families. A.T., E.A., and C.M. held fellowships from the FRIA (Belgium). COPPÉE F, MATTÉOTTI C, ANSEAU E, SAUVAGE S, LECLERCQ I, LEROY A, MARCOWYCZ A, GERBAUX C, FIGLEWICZ D, DING H, AND BELAYEW A. (2004) 117-134. Facioscapulohumeral Muscular Dystrophy: Clinical Medicine and Molecular Biology, M. Upadhyaya and D.N. Cooper, Eds, BIOS Scientific Publishers, Abingdon, UK. GABRIELS J., BECKERS M.C., DING H., DE VRIESE A., PLAISANCE S., VAN DER MAAREL S.M., PADBERG G.W., FRANTS R.R., HEWITT J.E., COLLEN D., AND BELAYEW A. (1999) Gene 236: 25-32. VAN DER MAAREL S.M., FRANTS R.R., PADBERG G.W. (2006) Biochim Biophys Acta. ePub Jun 6