Journal of Human Genetics

, Volume 52, Issue 4, pp 342–348

Mutation analysis of the MECP2 gene in patients of Slavic origin with Rett syndrome: novel mutations and polymorphisms

  • Daniela Zahorakova
  • Robert Rosipal
  • Jan Hadac
  • Alena Zumrova
  • Vladimir Bzduch
  • Nadezda Misovicova
  • Alice Baxova
  • Jiri Zeman
  • Pavel Martasek
Original Article

Abstract

Rett syndrome (RTT), an X-linked dominant neurodevelopmental disorder in females, is caused mainly by de novo mutations in the methyl-CpG-binding protein 2 gene (MECP2). Here we report mutation analysis of the MECP2 gene in 87 patients with RTT from the Czech and Slovak Republics, and Ukraine. The patients, all girls, with classical RTT were investigated for mutations using bi-directional DNA sequencing and conformation sensitive gel electrophoresis analysis of the coding sequence and exon/intron boundaries of the MECP2 gene. Restriction fragment length polymorphism analysis was performed to confirm the mutations that cause the creation or abolition of the restriction site. Mutation-negative cases were subsequently examined by multiple ligation-dependent probe amplification (MLPA) to identify large deletions. Mutation screening revealed 31 different mutations in 68 patients and 12 non-pathogenic polymorphisms. Six mutations have not been previously published: two point mutations (323T>A, 904C>T), three deletions (189_190delGA, 816_832del17, 1069delAGC) and one deletion/inversion (1063_1236del174;1189_1231inv43). MLPA analysis revealed large deletions in two patients. The detection rate was 78.16%. Our results confirm the high frequency of MECP2 mutations in females with RTT and provide data concerning the mutation heterogeneity in the Slavic population.

Keywords

Rett syndrome Mental retardation MECP2 gene Mutation screening MLPA 

Supplementary material

10038_2007_121_MOESM1_ESM.doc (38 kb)
MLPA probes in and around the MECP2 gene included in the SALSA MLPA P015C MECP2 Kit (according to the MRC Holland product sheet, http://www.mrc-holland.com/pages/p015pag.html) (DOC 37.5 KB)

References

  1. Amir RE, Zoghbi HY (2000) Rett syndrome: methyl-CpG-binding protein 2 mutations and phenotype–genotype correlations. Am J Med Genet 97:147–152PubMedCrossRefGoogle Scholar
  2. Amir RE, Van den Veyver IB, Wan M, Tran CQ, Francke U, Zoghbi HY (1999) Rett syndrome is caused by mutations in X-linked MECP2, encoding methyl-CpG-binding protein 2. Nat Genet 23:185–188PubMedCrossRefGoogle Scholar
  3. Amir RE, Fang P, Yu Z, Glaze DG, Percy AK, Zoghbi HY, Roa BB, Van den Veyver IB (2005) Mutations in exon 1 of MECP2 are a rare cause of Rett syndrome. J Med Genet 42:e15PubMedCrossRefGoogle Scholar
  4. Ballestar E, Yusufzai TM, Wolffe AP (2000) Effects of Rett syndrome mutations of the methyl-CpG binding domain of the transcriptional repressor MeCP2 on selectivity for association with methylated DNA. Biochemistry 39:7100–7106PubMedCrossRefGoogle Scholar
  5. Borg I, Freude K, Kubart S, Hoffmann K, Menzel C, Laccone F, Firth H, Ferguson-Smith MA, Tommerup N, Ropers HH, Sargan D, Kalscheuer VM (2005) Disruption of Negrin G1 by a balanced chromosome translocation in a girl with Rett syndrome. Eur J Hum Genet 13:921–927PubMedCrossRefGoogle Scholar
  6. Bourdon V, Philippe Ch, Labrune O, Amsallem D, Arnould C, Jonveaux P (2001) A detailed analysis of the MECP2 gene: prevalence of recurrent mutations and gross DNA rearrangements in Rett syndrome patients. Hum Genet 108:43–50PubMedCrossRefGoogle Scholar
  7. Buyse IM, Fang P, Hoon KT, Amir RE, Zoghbi HY, Roa BB (2000) Diagnostic testing for Rett syndrome by DHPLC and direct sequencing analysis of the MECP2 gene: identification of several novel mutations and polymorphisms. Am J Hum Genet 67:1428–1436PubMedCrossRefGoogle Scholar
  8. Cheadle JP, Gill H, Fleming N, Maynard J, Kerr A, Leonard H, Krawczak M, Cooper DN, Lynch S, Thomas N, Hughes H, Hulten M, Ravine D, Sampson JR, Clarke A (2000) Long-read sequence analysis of the MECP2 gene in Rett syndrome patients: correlation of disease severity with mutation type and location. Hum Mol Genet 9:1119–1129PubMedCrossRefGoogle Scholar
  9. Conforti FL, Mazzei R, Magariello A, Patitucci A, Gabriele AL, Muglia M, Quattrone A, Fiumara A, Barone R, Pavone L, Nistico R, Mangone L (2003) Mutation analysis of the MECP2 gene in patients with Rett syndrome. Am J Med Genet 117A:184–187CrossRefGoogle Scholar
  10. Couvert P, Bienvenu T, Aquaviva C, Poirier K, Moraine C, Gendrot C, Verloes A, Andres C, Le Fevre AC, Souville I, Steffann J, des Portes V, Ropers HH, Yntema HG, Fryns JP, Briault S, Chelly J, Cherif B (2001) MECP2 is highly mutated in X-linked mental retardation. Hum Mol Genet 10:941–946PubMedCrossRefGoogle Scholar
  11. De Bona C, Zappella M, Hayek G, Meloni I, Vitelli F, Bruttini M, Cusano R, Loffredo P, Longo I, Renieri A (2000) Preserved speech variant is allelic of classic Rett syndrome. Eur J Hum Genet 8:325–330PubMedCrossRefGoogle Scholar
  12. Erlandson A, Hallberg B, Hagberg B, Wahlstron J, Martinsson T (2001) MECP2 mutation screening in Swedish classical Rett syndrome females. Eur Child Adolesc Psychiatry 10:117–121PubMedCrossRefGoogle Scholar
  13. Evans JC, Archer H, Whatley SD, Kerr A, Clarke A, Butler R (2005) Variation in exon 1 coding region and promoter of MECP2 in Rett syndrome and controls. Eur J Hum Genet 13:124–126PubMedCrossRefGoogle Scholar
  14. Hagberg B (1995) Clinical delineation of Rett syndrome variants. Neuropediatrics 26:62PubMedCrossRefGoogle Scholar
  15. Hagberg B, Aicardi J, Dias K, Ramos O (1983) A progressive syndrome of autism, dementia, ataxia, and loss of purposeful hand use in girls: Rett’s syndrome: report of 35 cases. Ann Neurol 14:471–479PubMedCrossRefGoogle Scholar
  16. Hoffbuhr K, Devaney JM, LaFleur B, Sirianni N, Scacheri C, Giron J, Schuette J, Innis J, Marino M, Philippart M, Narayanan V, Umansky R, Kronn D, Hoffman EP, Naidu S (2001) MeCP2 mutations in children with and without the phenotype of Rett syndrome. Neurology 56:1486–1495PubMedGoogle Scholar
  17. Hoffbuhr KC, Moses LM, Jerdonek MA, Naidu S, Hoffman EP (2002) Associations between MECP2 mutations, X-chromosome inactivation, and phenotype. Ment Retard Dev Disabil Res Rev 8:99–105PubMedCrossRefGoogle Scholar
  18. Huppke P, Laccone F, Kramer N, Engel W, Hanefeld F (2000) Rett syndrome: analysis of MECP2 and clinical characterization of 31 patients. Hum Mol Genet 9:1369–1375PubMedCrossRefGoogle Scholar
  19. Jones PL, Veenstra GJ, Wade PA, Wade PA, Vermaak D, Kass SU, Landsberger N, Strouboulis J, Wolffe AP (1998) Methylated DNA and MeCP2 recruit histone deacetylase to repress transcription. Nat Genet 19:187–191PubMedCrossRefGoogle Scholar
  20. Kerr AM, Nomura Y, Armstrong D, Anvret M, Belischenko PV, Budden S, Cass H, Christodoulou J, Clarke A, Ellaway C, d’Esposito M, Francke U, Hulten M, Julu P, Leonard H, Naidu S, Schanen C, Webb T, Engerstrom IW, Yamashita Y, Segawa M (2001) Guidelines for reporting clinical features in cases with MECP2 mutations. Brain Dev 23:208–211PubMedCrossRefGoogle Scholar
  21. Laccone F, Huppke P, Hanefeld F, Meins M (2001) Mutation spectrum in patients with Rett syndrome in the German population: evidence of hot spot regions. Hum Mutat 17:183–190PubMedCrossRefGoogle Scholar
  22. Laccone F, Junemann I, Whatley S, Morgan R, Butler R, Huppke P, Ravine D (2004) Large deletions of the MECP2 gene detected by gene dosage analysis in patients with Rett syndrome. Hum Mutat 23:234–244PubMedCrossRefGoogle Scholar
  23. Mari F, Azimonti S, Bertani I, Bolognese F, Colombo E, Caselli R, Scala E, Longo I, Grosso S, Pescucci C, Ariani F, Hayek G, Balestri P, Bergo A, Badaracco G, Zapella M, Broccoli V, Renieri A, Kilstrup-Nielsen C, Landsberger N (2005) CDKL5 belongs to the same molecular pathway of MeCP2 and it is responsible for the early-onset seizure variant of Rett syndrome. Hum Mol Genet 14:1935–1946PubMedCrossRefGoogle Scholar
  24. Mnatzakanian GN, Lohi H, Munteanu I, Alfred SE, Yamada T, MacLeod PJ, Jones JR, Scherer SW, Schanen NC, Friez MJ, Vincent JB, Minassian BA (2004) A previously unidentified MECP2 open reading frame defines a new protein isoform relevant to Rett syndrome. Nat Genet 36:339–341PubMedCrossRefGoogle Scholar
  25. Naidu S (1997) Rett syndrome: natural history and underlying disease mechanisms. Eur Child Adolesc Psychiatry 6(Suppl 1):14–17PubMedGoogle Scholar
  26. Nan X, Ng HH, Johnson CA, Laherty CD, Turner BM, Eisenman RN, Bird A (1998) Transcriptional repression by the methyl-CpG-binding protein MeCP2 involves a histone deacetylase complex. Nature 393:386–389PubMedCrossRefGoogle Scholar
  27. Rett A (1966) Über ein eigenartiges hirnatrophisches Syndrom bei Hyperammonemie im Kindesalter. Wien Med Wochenschr 116:723–738PubMedGoogle Scholar
  28. Shibayama A, Cook EH, Feng J, Glanzmann C, Yan J, Craddock N, Jones IR, Goldman D, Heston LL, Sommer SS (2004) MECP2 structural and 3′-UTR variants in schizophrenia, autism and other psychiatric diseases: a possible association with autism. Am J Med Genet Part B (Neuropsychiatr Genet) 128:50–53CrossRefGoogle Scholar
  29. Tao J, Van Esch H, Hegedorn-Greiwr M, Hoffmann K, Moser B, Raynaud M, Sperner J, Fryns JP, Schwinger E, Gecz J, Ropers HH, Kalscheuer VM (2004) Mutations in the X-linked cyclin-dependent kinase-like 5 (CDKL5/STK9) gene are associated with severe neurodevelopmental retardation. Am J Hum Genet 75:1149–1154PubMedCrossRefGoogle Scholar
  30. Van den Veyver IB, Zoghbi HY (2000) Methyl-CpG-binding protein 2 mutations in Rett syndrome. Curr Opin Genet Dev 10:275–279PubMedCrossRefGoogle Scholar
  31. Wan M, Lee SS, Zhang X, Houwink-Manville I, Song HR, Amir RE, Budden S, Naidu S, Pereira JL, Lo IF, Zoghbi HY, Schanen NC, Francke U (1999) Rett syndrome and beyond: recurrent spontaneous and familial MECP2 mutations at CpG hotspots. Am J Hum Genet 65:1520–1529PubMedCrossRefGoogle Scholar
  32. Weaving LS, Christodoulou J, Williamson SL, Friend KL, McKenzie OL, Archer H, Evans J, Clarke A, Pelka GJ, Tam PP, Watson C, Lahooti H, Ellaway CJ, Bennetts B, Leonard H, Gecz J (2004) Mutations of CDKL5 cause a severe neurodevelopmental disorder with infantile spasms and mental retardation. Am J Hum Genet 75:1079–1093PubMedCrossRefGoogle Scholar
  33. Young JI, Hong EP, Castle JC, Crespo-Barreto J, Bowman AB, Rose MF, Kang D, Richman R, Johnson JM, Berget S, Zoghbi HY (2004) Regulation of RNA splicing by the methylation-dependent transcriptional repressor methyl-CpG binding protein 2. Proc Natl Acad Sci USA 102:17551–17558CrossRefGoogle Scholar
  34. Zeev BB, Yaron Y, Schanen NC, Wolf H, Brandt N, Ginot N, Shomrat R, Orr-Urtreger A (2002) Rett syndrome: clinical manifestations in males with MECP2 mutations. J Child Neurol 17:20–24PubMedGoogle Scholar

Copyright information

© The Japan Society of Human Genetics and Springer 2007

Authors and Affiliations

  • Daniela Zahorakova
    • 1
  • Robert Rosipal
    • 1
  • Jan Hadac
    • 2
  • Alena Zumrova
    • 3
  • Vladimir Bzduch
    • 4
  • Nadezda Misovicova
    • 5
  • Alice Baxova
    • 6
  • Jiri Zeman
    • 1
  • Pavel Martasek
    • 1
  1. 1.Department of Pediatrics, First School of MedicineCharles UniversityPrague 2Czech Republic
  2. 2.Department of Child NeurologyThomayer University HospitalPragueCzech Republic
  3. 3.Department of Child NeurologyUniversity Hospital MotolPragueCzech Republic
  4. 4.First Department of PediatricsCommenius University Children’s HospitalBratislavaSlovakia
  5. 5.Department of Clinical GeneticsMartin University HospitalMartinSlovakia
  6. 6.Institute of Biology and Clinical GeneticsGeneral University HospitalPragueCzech Republic

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