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Methylation status of ribosomal RNA gene clusters in the flow-sorted human acrocentric chromosomes

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Abstract

Southern blot analysis of the human acrocentric chromosomes that were flow-sorted from B-lymphoblastoid cell line GM130B revealed that the sensitivity of the ribosomal RNA (rDNA) gene clusters to the restriction enzyme NotI differs among these rDNA-containing chromosomes: the rDNA clusters of Chromosomes (Chr) 13, 14, and 15 are much more sensitive to NotI digestion than those of Chrs 21 and 22 in this particular cell line. Detailed analysis by use of methylation-sensitive enzymes HpaII and HhaI and methylation-insensitive enzyme MspI confirmed the significant variation in the methylation status of rDNA clusters among these chromosomes. Quantitative analysis by fluorescent in situ hybridization (FISH) indicated that copy number of rDNA varies among individual chromosomes, but the average copy number in the acrocentric Chrs 21 and 22 is significantly greater than that of the Chrs 13, 14, and 15 in GM130B cells. Similar analysis reveals that the methylation status of rDNA clusters in another B-lymphoblastoid cell line GM131 was different from that of GM130B. These data together indicate that the copy number and methylation patterns of rDNA clusters differ among individual acrocentric chromosomes in a given cell line, and they are different among cell lines.

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References

  • Bernstein, R., Dawson, B., and Griffiths, J.: Human inherited marker Chromosome 22 short-arm enlargement: investigation of rDNA gene multiplicity, Ag-band size, and acrocentric association. Hum Genet 58: 135–139, 1981.

    Google Scholar 

  • Bird, A.P. and Taggart, M.H.: Variable patterns of total DNA and rDNA methylation in animals. Nucleic Acids Res 8: 1485–1497, 1980.

    Google Scholar 

  • Bird, A.P., Taggart, M.H., and Gehring, C.A.: Methylated and unmethylated ribosomal RNA genes in the mouse. J Mol Biol 152: 1–17, 1981.

    Google Scholar 

  • de Capoa, A., Felli, M.P., Baldini, A., Rocci, M., Archidiacono, N., Aleixandre, C., Miller, O.J., and Miller, D.A.: Relationship between the number and function of human ribosomal genes. Hum Genet 79: 301–304, 1988.

    Google Scholar 

  • Feinberg, A.P., and Vogelstein, B.: A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13, 1983.

    Google Scholar 

  • Ferguson-Smith, M.A. and Handmarker, S.D.: Observations on the satellited human chromosomes. Lancet 1: 638–640, 1961.

    Google Scholar 

  • Ferraro, M. and Lavia, P.: Activation of human ribosomal genes by 5-azacytidine. Exp Cell Res 145: 452–457, 1983.

    Google Scholar 

  • Gonzalez, I.L., Gorski, J.L., Campen, T.J., Dorney, D.J., Erickson, J.M., Sylvester, J.E., and Schmickel, R.D.: Variation among human 28S ribosomal RNA genes. Proc Natl Acad Sci USA 82: 7666–7670, 1985.

    Google Scholar 

  • Goodpasture, C. and Bloom, S.E.: Visualization of nucleolus organizer regions in mammalian chromosomes using silver stain. Chromosoma 53: 37–50, 1975.

    Google Scholar 

  • Henderson, A.S. and Atwood, K.C.: Satellite-association frequency and rDNA content of a double-satellited chromosome. Hum Genet 31: 113–115, 1976.

    Google Scholar 

  • Henderson, A.S., Warburton, D., and Atwood, K.C.: Location of ribosomal DNA in the human chromosome complement. Proc Natl Acad Sci USA 69: 3394–3398, 1972.

    Google Scholar 

  • Kawasaki, K., Kudoh, J., Omoto, K., and Shimizu, N.: Mega base map of the epidermal growth factor (EGF) receptor gene flanking regions and structure of the amplification units in EGF receptor-hyperproducing squamous carcinoma cells. Jpn J Cancer Res (Gann) 79: 1174–1183, 1988.

    Google Scholar 

  • Krystal, M., D'Eustachio, P., Ruddle, F.H., Arnheim, N.: Human nucleolus organizers on nonhomologous chromosomes can share the same ribosomal gene variants. Proc Natl Acad Sci USA 78: 5744–5748, 1981.

    Google Scholar 

  • Lau, Y.-F., Wertelecki, W., Pfeiffer, R.A., and Arrighi, F.E.: Cytological analyses of a 14p+ variant by means of N-banding and combinations of silver staining and chromosome bandings. Hum Genet 46: 75–82, 1979.

    Google Scholar 

  • Lindsay, S. and Bird, A.P.: use of restriction enzymes to detect potential gene sequences in mammalian DNA. Nature 327: 336–338, 1987.

    Google Scholar 

  • Miller, D.A., Tantravahi, R., Dev, V.G., and Miller, O.J.: Frequency of satellite association of human chromosomes is correlated with amount of Ag-staining of the nucleolus organizer region. Am J Hum Genet 29: 490–502, 1977.

    Google Scholar 

  • Miller, D.A., Roy Breg, W., Warturton, D., Dev, V.G., and Miller, O.J.: Regulation of rRNA gene expression in a human familial 14p+ marker chromosome. Hum Genet 43: 289–297, 1978.

    Google Scholar 

  • Miller, O.J., Miller, D.A., Dev, V.G., Tantravahi, R., and Croce, C.M.: Expression of human and suppression of mouse nucleolus organizer activity in mouse-human somatic cell hybrids. Proc Natl Acad Sci USA 73: 4531–4535, 1976.

    Google Scholar 

  • Minoshima, S., Kawasaki, K., Fukuyama, R., Maekawa, M., Kudoh, J., and Shimizu, N.: Isolation of giant DNA fragments from flow-sorted human chromosomes. Cytometry 11: 539–546, 1990.

    Google Scholar 

  • Nelson, M. and McClelland, M.: Effect of site-specific methylation on DNA modification methyltransferases and restriction endonucleases. Nucleic Acids Res 17: r389-r415, 1989.

    Google Scholar 

  • Pinkel, D., Straume, T., and Gray, J.M.: Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridization. Proc Natl Acad Sci USA 83: 2934–2938, 1986.

    Google Scholar 

  • Razin, A. and Riggs, A.D.: DNA methylation and gene function. Science 210: 604–610, 1980.

    Google Scholar 

  • Reilly, J.G., Thomas, C.A. Jr, and Lundell, M.J.: Methylation of mouse ribosomal RNA genes. DNA 1: 259–266, 1982.

    Google Scholar 

  • Sakai, K., Hirai, M., Minoshima, S., Kudoh, J., Fukuyama, R., and Shimizu, N.: Isolation of cDNAs encoding a substrate for protein kinase C: nucleotide sequence and chromosomal mapping of the gene for a human 80K protein. Genomics 5: 309–315, 1989.

    Google Scholar 

  • Tantravahi, U., Guntaka, R.V., Erlanger B.F., and Miller, O.J.: Amplified ribosomal RNA genes in a rat hepatoma cell line are enriched in 5-methylcytosine. Proc Natl Acad Sci USA 78: 489–492, 1981.

    Google Scholar 

  • Warburton, D. and Henderson, A.S.: Sequential silver staining and hybridization in situ on nucleolus organizing regions in human cells. Cytogenet Cell Genet 24: 168–175, 1979.

    Google Scholar 

  • Warburton, D., Atwood, K.C., and Henderson, A.S.: Variation in the number of genes for rRNA among human acrocentric chromosomes: correlation with frequency of satellite association. Cytogenet Cell Genet 17: 221–230, 1976.

    Google Scholar 

  • White, J.G., Amos, W.B., and Fordham, M.: An evaluation of confocal versus conventional imaging of biological structures by fluorescence light microscopy. J Cell Biol 105: 41–48, 1987.

    Google Scholar 

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Kawasaki, K., Minoshima, S., Kudoh, J. et al. Methylation status of ribosomal RNA gene clusters in the flow-sorted human acrocentric chromosomes. Mammalian Genome 3, 173–178 (1992). https://doi.org/10.1007/BF00352463

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