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. 1997 Feb;17(2):707–712. doi: 10.1128/mcb.17.2.707

A unique downregulation of h2-calponin gene expression in Down syndrome: a possible attenuation mechanism for fetal survival by methylation at the CpG island in the trisomic chromosome 21.

J Kuromitsu 1, H Yamashita 1, H Kataoka 1, T Takahara 1, M Muramatsu 1, T Sekine 1, N Okamoto 1, Y Furuichi 1, Y Hayashizaki 1
PMCID: PMC231796  PMID: 9001224

Abstract

To understand the effect of trisomic chromosome 21 on the cause of Down syndrome (DS), DNA methylation in the CpG island, which regulates the expression of adjacent genes, was investigated with the DNAs of chromosome 21 isolated from DS patients and their parents. A methylation-sensitive enzyme, BssHII, was used to digest DNAs of chromosome 21, and the resulting DNA fragments were subjected to RLGS (restriction landmark genomic scanning). Surprisingly, the CpG island of the h2-calponin gene was shown to be specifically methylated by comparative studies with RLGS and Southern blot analysis. In association with this methylation, h2-calponin gene expression was attenuated to the normal level, although other genes in the DS region of chromosome 21 were expressed dose dependently at 1.5 times the normal level. These results and the high miscarriage rate associated with trisomy 21 embryos imply that the altered in vivo methylation that attenuates downstream gene expression, which is otherwise lethal, permits the generation of DS neonates. The h2-calponin gene detected by the RLGS procedure may be one such gene that is attenuated.

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Selected References

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  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Bird A. The essentials of DNA methylation. Cell. 1992 Jul 10;70(1):5–8. doi: 10.1016/0092-8674(92)90526-i. [DOI] [PubMed] [Google Scholar]
  3. Cedar H. DNA methylation and gene activity. Cell. 1988 Apr 8;53(1):3–4. doi: 10.1016/0092-8674(88)90479-5. [DOI] [PubMed] [Google Scholar]
  4. Cheng J. F., Boyartchuk V., Zhu Y. Isolation and mapping of human chromosome 21 cDNA: progress in constructing a chromosome 21 expression map. Genomics. 1994 Sep 1;23(1):75–84. doi: 10.1006/geno.1994.1461. [DOI] [PubMed] [Google Scholar]
  5. Coyle J. T., Oster-Granite M. L., Gearhart J. D. The neurobiologic consequences of Down syndrome. Brain Res Bull. 1986 Jun;16(6):773–787. doi: 10.1016/0361-9230(86)90074-2. [DOI] [PubMed] [Google Scholar]
  6. Frischer H., Chu L. K., Ahmad T., Justice P., Smith G. F. Superoxide dismutase and glutathione peroxidase abnormalities in erythrocytes and lymphoid cells in Down syndrome. Prog Clin Biol Res. 1981;55:269–289. [PubMed] [Google Scholar]
  7. Gearhart J. D., Davisson M. T., Oster-Granite M. L. Autosomal aneuploidy in mice: generation and developmental consequences. Brain Res Bull. 1986 Jun;16(6):789–801. doi: 10.1016/0361-9230(86)90075-4. [DOI] [PubMed] [Google Scholar]
  8. Gerdes A. M., Hørder M., Petersen P. H., Bonnevie-Nielsen V. Effect of increased gene dosage expression on the alpha-interferon receptors in Down's syndrome. Biochim Biophys Acta. 1993 Apr 30;1181(2):135–140. doi: 10.1016/0925-4439(93)90102-7. [DOI] [PubMed] [Google Scholar]
  9. Hayashizaki Y., Hirotsune S., Okazaki Y., Hatada I., Shibata H., Kawai J., Hirose K., Watanabe S., Fushiki S., Wada S. Restriction landmark genomic scanning method and its various applications. Electrophoresis. 1993 Apr;14(4):251–258. doi: 10.1002/elps.1150140145. [DOI] [PubMed] [Google Scholar]
  10. Hayashizaki Y., Shibata H., Hirotsune S., Sugino H., Okazaki Y., Sasaki N., Hirose K., Imoto H., Okuizumi H., Muramatsu M. Identification of an imprinted U2af binding protein related sequence on mouse chromosome 11 using the RLGS method. Nat Genet. 1994 Jan;6(1):33–40. doi: 10.1038/ng0194-33. [DOI] [PubMed] [Google Scholar]
  11. Hirotsune S., Shibata H., Okazaki Y., Sugino H., Imoto H., Sasaki N., Hirose K., Okuizumi H., Muramatsu M., Plass C. Molecular cloning of polymorphic markers on RLGS gel using the spot target cloning method. Biochem Biophys Res Commun. 1993 Aug 16;194(3):1406–1412. doi: 10.1006/bbrc.1993.1981. [DOI] [PubMed] [Google Scholar]
  12. Holtzman D. M., Epstein C. J. The molecular genetics of Down syndrome. Mol Genet Med. 1992;2:105–120. doi: 10.1016/b978-0-12-462002-5.50009-1. [DOI] [PubMed] [Google Scholar]
  13. Kato K., Suzuki F., Kurobe N., Okajima K., Ogasawara N., Nagaya M., Yamanaka T. Enhancement of S-100 beta protein in blood of patients with Down's syndrome. J Mol Neurosci. 1990;2(2):109–113. doi: 10.1007/BF02876918. [DOI] [PubMed] [Google Scholar]
  14. Korenberg J. R., Chen X. N., Schipper R., Sun Z., Gonsky R., Gerwehr S., Carpenter N., Daumer C., Dignan P., Disteche C. Down syndrome phenotypes: the consequences of chromosomal imbalance. Proc Natl Acad Sci U S A. 1994 May 24;91(11):4997–5001. doi: 10.1073/pnas.91.11.4997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kuromitsu J., Kataoka H., Yamashita H., Muramatsu M., Furuichi Y., Sekine T., Hayashizaki Y. Reproducible alterations of DNA methylation at a specific population of CpG islands during blast formation of peripheral blood lymphocytes. DNA Res. 1995 Dec 31;2(6):263–267. doi: 10.1093/dnares/2.6.263. [DOI] [PubMed] [Google Scholar]
  16. Langlois R. G., Yu L. C., Gray J. W., Carrano A. V. Quantitative karyotyping of human chromosomes by dual beam flow cytometry. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7876–7880. doi: 10.1073/pnas.79.24.7876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lemieux N., Malfoy B., Forrest G. L. Human carbonyl reductase (CBR) localized to band 21q22.1 by high-resolution fluorescence in situ hybridization displays gene dosage effects in trisomy 21 cells. Genomics. 1993 Jan;15(1):169–172. doi: 10.1006/geno.1993.1024. [DOI] [PubMed] [Google Scholar]
  18. Lindsay S., Bird A. P. Use of restriction enzymes to detect potential gene sequences in mammalian DNA. 1987 May 28-Jun 3Nature. 327(6120):336–338. doi: 10.1038/327336a0. [DOI] [PubMed] [Google Scholar]
  19. Masuda H., Tanaka K., Takagi M., Ohgami K., Sakamaki T., Shibata N., Takahashi K. Molecular cloning and characterization of human non-smooth muscle calponin. J Biochem. 1996 Aug;120(2):415–424. doi: 10.1093/oxfordjournals.jbchem.a021428. [DOI] [PubMed] [Google Scholar]
  20. Nakajima H., Kono N., Yamasaki T., Hamaguchi T., Hotta K., Kuwajima M., Noguchi T., Tanaka T., Tarui S. Tissue specificity in expression and alternative RNA splicing of human phosphofructokinase-M and -L genes. Biochem Biophys Res Commun. 1990 Dec 31;173(3):1317–1321. doi: 10.1016/s0006-291x(05)80931-3. [DOI] [PubMed] [Google Scholar]
  21. Ohsumi T., Okazaki Y., Hirotsune S., Shibata H., Muramatsu M., Suzuki H., Taga C., Watanabe S., Hayashizaki Y. A spot cloning method for restriction landmark genomic scanning. Electrophoresis. 1995 Feb;16(2):203–209. doi: 10.1002/elps.1150160135. [DOI] [PubMed] [Google Scholar]
  22. Okazaki Y., Okuizumi H., Sasaki N., Ohsumi T., Kuromitsu J., Kataoka H., Muramatsu M., Iwadate A., Hirota N., Kitajima M. A genetic linkage map of the mouse using an expanded production system of restriction landmark genomic scanning (RLGS Ver.1.8). Biochem Biophys Res Commun. 1994 Dec 30;205(3):1922–1929. doi: 10.1006/bbrc.1994.2895. [DOI] [PubMed] [Google Scholar]
  23. Pash J., Smithgall T., Bustin M. Chromosomal protein HMG-14 is overexpressed in Down syndrome. Exp Cell Res. 1991 Mar;193(1):232–235. doi: 10.1016/0014-4827(91)90562-9. [DOI] [PubMed] [Google Scholar]
  24. Strasser P., Gimona M., Moessler H., Herzog M., Small J. V. Mammalian calponin. Identification and expression of genetic variants. FEBS Lett. 1993 Sep 6;330(1):13–18. doi: 10.1016/0014-5793(93)80909-e. [DOI] [PubMed] [Google Scholar]
  25. Suzuki H., Kawai J., Taga C., Ozawa N., Watanabe S. A PCR-mediated method for cloning spot DNA on restriction landmark genomic scanning (RLGS) gel. DNA Res. 1994;1(5):245–250. doi: 10.1093/dnares/1.5.245. [DOI] [PubMed] [Google Scholar]
  26. Takahashi K., Nadal-Ginard B. Molecular cloning and sequence analysis of smooth muscle calponin. J Biol Chem. 1991 Jul 15;266(20):13284–13288. [PubMed] [Google Scholar]
  27. Taylor G. M., Williams A., D'Souza S. W., Fergusson W. D., Donnai D., Fennell J., Harris R. The expression of CD18 is increased on Trisomy 21 (Down syndrome) lymphoblastoid cells. Clin Exp Immunol. 1988 Feb;71(2):324–328. [PMC free article] [PubMed] [Google Scholar]
  28. Vora S., Francke U. Assignment of the human gene for liver-type 6-phosphofructokinase isozyme (PFKL) to chromosome 21 by using somatic cell hybrids and monoclonal anti-L antibody. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3738–3742. doi: 10.1073/pnas.78.6.3738. [DOI] [PMC free article] [PubMed] [Google Scholar]

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