Myotonic Dystrophy Type 2 Caused by a CCTG Expansion in Intron 1 of ZNF9
10.1126/science.1062125
Crossref journal-article
American Association for the Advancement of Science (AAAS)
Science (221)
Abstract

Myotonic dystrophy (DM), the most common form of muscular dystrophy in adults, can be caused by a mutation on either chromosome 19q13 (DM1) or 3q21 (DM2/PROMM). DM1 is caused by a CTG expansion in the 3′ untranslated region of the dystrophia myotonica–protein kinase gene ( DMPK ). Several mechanisms have been invoked to explain how this mutation, which does not alter the protein-coding portion of a gene, causes the specific constellation of clinical features characteristic of DM. We now report that DM2 is caused by a CCTG expansion (mean ∼5000 repeats) located in intron 1 of the zinc finger protein 9 ( ZNF9 ) gene. Parallels between these mutations indicate that microsatellite expansions in RNA can be pathogenic and cause the multisystemic features of DM1 and DM2.

Bibliography

Liquori, C. L., Ricker, K., Moseley, M. L., Jacobsen, J. F., Kress, W., Naylor, S. L., Day, J. W., & Ranum, L. P. W. (2001). Myotonic Dystrophy Type 2 Caused by a CCTG Expansion in Intron 1 of ZNF9. Science, 293(5531), 864–867.

Authors 8
  1. Christina L. Liquori (first)
  2. Kenneth Ricker (additional)
  3. Melinda L. Moseley (additional)
  4. Jennifer F. Jacobsen (additional)
  5. Wolfram Kress (additional)
  6. Susan L. Naylor (additional)
  7. John W. Day (additional)
  8. Laura P. W. Ranum (additional)
References 49 Referenced 1,012
  1. P. S. Harper Myotonic Dystrophy (Saunders London ed. 2 1989).
  2. 10.1007/BF01671719
  3. 10.1002/ana.410350305
  4. 10.1212/WNL.44.8.1448
  5. 10.1002/(SICI)1521-1878(199811)20:11<901::AID-BIES5>3.0.CO;2-0
  6. 10.1126/science.289.5485.1701
  7. 10.1126/science.8469976
  8. 10.1093/hmg/4.10.1919
  9. 10.1073/pnas.92.12.5465
  10. 10.1038/ng0897-402
  11. 10.1038/ng0897-407
  12. 10.1093/hmg/8.8.1491
  13. 10.1083/jcb.128.6.995
  14. 10.1073/pnas.94.14.7388
  15. 10.1093/nar/24.22.4407
  16. 10.1126/science.280.5364.737
  17. 10.1093/hmg/8.1.53
  18. 10.1093/emboj/19.17.4439
  19. 10.1126/science.289.5485.1769
  20. 10.1038/ng0796-325
  21. 10.1038/75490
  22. 10.1038/75500
  23. 10.1038/570
  24. 10.1016/S0960-8966(98)00094-7
  25. We identified obtained informed consent performed neurological exams and collected blood samples from DM2/PROMM family members. Genomic DNA was isolated from blood using the Puregene kit #D-5000 (Gentra Systems Minneapolis MN). Linkage analysis was performed using the LINKAGE package of computer programs (version 5.1) (48).
  26. 10.1038/35057157
  27. Supplemental data are available on Science Online at www.sciencemag.org/cgi/content/full/293/5531/864/DC1
  28. Spielman R. S., McGinnis R. E., Ewens W. J., Am. J. Hum. Genet. 52, 506 (1993). / Am. J. Hum. Genet. by Spielman R. S. (1993)
  29. We amplified the DM2 repeat region from genomic DNA using primers CL3N58-D F (5′-GCCTAGGGGACAAAGTGAGA-3′) and CL3N58-D R (5′-GGCCTTATAACCATGCAAATG-3′) in a PCR reaction [200 μM deoxynucleotide triphosphates (dNTPs) 10 mM tris-HCl (pH 9.0) 50 mM KCl 0.1% Triton X-100 0.01% (w/v) gelatin 1 mM MgCl 2 0.4 μM each primer 0.1 U Taq] cycled 30 times (94°C for 45 s 57°C for 45 s 72°C for 1 min). Unrelated control DNA samples included the grandparents from the panel of 40 Centre d'Etude du Polymorphisme Humain (CEPH) families spouses of patients diagnosed with muscular dystrophy or ataxia and ataxia patients ( n = 1360 chromosomes). Small expanded alleles were amplified from genomic DNA using primers CL3N58-B F (5′-TGAGCCGGAATCATACCAGT-3′) and CL3N58-D R in a PCR reaction [200 μM dNTPs 50 mM Tris-HCl (pH 9.1) 14 mM (NH 4 )SO 4 2 mM MgCl 2 0.4 μM each primer 0.1% Tween-20 10% dimethyl sulfoxide 0.75 U ProofSprinter enzyme (Hybaid-AGS)] cycled 35 times (94°C for 30 s 51°C for 30 s 72°C for 1 min). Normal and expanded alleles amplified by PCR were cloned with the TOPO cloning kit (Invitrogen Carlsbad CA) and sequenced.
  30. Bso BI–digested genomic DNA (5 μg) was separated on an 0.8% agarose gel transferred to Hybond N+ membrane (Amersham Piscataway NJ) and hybridized with a 474-bp ZNF9 probe using Rapid-Hyb buffer (Amersham). The probe was generated by PCR using the primers probeA F (5′-GAGAACCTTGCCATTTTTCG-3′) and probeA R (5′-CACCTACAGCACTGGCAACA-3′) and random-prime-labeled (GibcoBRL Carlsbad CA) with 32 P-α-deoxyadenosine triphosphate (NEN Boston MA). To avoid partial digestions with Bso BI we used 120 U of enzyme in a digestion volume of 120 μl. For more accurate sizing of the high molecular weight expansions Eco RI–digested genomic DNA (5 μg) was separated on a 0.4% agarose gel along with high molecular weight DNA markers (Gibco-BRL).
  31. L. P. W. Ranum J. W. Day unpublished data.
  32. GenBank accession numbers are as follows: genomic sequence of the DM2 region ( ); CL3N58 sequence (); expanded CL3N58 sequence (); ZNF9 mRNA (); original ZNF9 genomic sequence (); RP11-814L21 (); RP11-723o4 (); and RP11-221e20 (). The Celera accession number for the contig overlapping ZNF9 is x2HTBKUAD8C.
  33. 10.1126/science.1058040
  34. 10.1126/science.2562787
  35. 10.1006/jmbi.1996.0888
  36. 10.1006/jmbi.1998.1961
  37. 10.1016/0378-1119(95)00421-2
  38. For in situ hybridization of muscle sections (13) we used 0.2 ng/μl 2′-O-methyl RNA oligos 5′ labeled with Cy3 (IDT Coralville IA). Fluorescence was visualized using a Zeiss Axioplan2 microscope equipped with a Spot CCD (charge-coupled device) camera (Diagnostic Instruments Sterling Heights MI). Appropriate exposure times were computed using the DM2/CAGG slide and the other probes were photographed using this exposure setting.
  39. 10.1006/bbrc.2000.3694
  40. 10.1038/7710
  41. 10.1038/79911
  42. Wong L. J., Ashizawa T., Monckton D. G., Caskey C. T., Richards C. S., Am. J. Hum. Genet. 56, 114 (1995). / Am. J. Hum. Genet. by Wong L. J. (1995)
  43. 10.1093/hmg/6.7.971
  44. A. T. Helderman-van den Enden et al. J. Med. Genet. 36 253 (1999).
  45. 10.1002/ana.410350323
  46. 10.1212/WNL.55.3.383
  47. 10.1212/WNL.52.1.170
  48. 10.1073/pnas.81.11.3443
  49. We thank families and clinicians for their participation K. Dick and L. Rasmussen for technical assistance J. Dalton and A. Dudley for organizing family evaluations K. Gebhard and J. Sedgwick for microphotography M. C. Koch and D. Garcia for genetic analysis and L. Ptacek for providing DNA from a family. Supported by the Muscle Center Clinical Research Center and Graduate School at the University of Minnesota; Förderverein of the Department of Neurology University of Würzburg; Research Funds from the State of Bavaria; the Muscular Dystrophy Association USA; and NIH grants NS35870 HG002051 and CA56266.
Dates
Type When
Created 23 years, 1 month ago (July 27, 2002, 5:39 a.m.)
Deposited 1 year, 7 months ago (Jan. 9, 2024, 5:11 p.m.)
Indexed 3 days, 10 hours ago (Aug. 26, 2025, 2:49 a.m.)
Issued 24 years ago (Aug. 3, 2001)
Published 24 years ago (Aug. 3, 2001)
Published Print 24 years ago (Aug. 3, 2001)
Funders 0

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@article{Liquori_2001, title={Myotonic Dystrophy Type 2 Caused by a CCTG Expansion in Intron 1 of ZNF9}, volume={293}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.1062125}, DOI={10.1126/science.1062125}, number={5531}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Liquori, Christina L. and Ricker, Kenneth and Moseley, Melinda L. and Jacobsen, Jennifer F. and Kress, Wolfram and Naylor, Susan L. and Day, John W. and Ranum, Laura P. W.}, year={2001}, month=aug, pages={864–867} }