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Restoration of the cholesterol metabolism in 3T3 cell lines derived from the sphingomyelinosis mouse (spm/spm) by transfer of a human chromosome 18

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Abstract

We searched for a human chromosome that would restore the cholesterol metabolism in 3T3 cell lines (SPM-3T3) derived from homozygous sphingomyelinosis mice (spm/spm). Mouse A9 cells containing a single copy of pSV2neo-tagged chromosomes 9, 11, or 18 derived from normal human fibroblasts served as donor cells for transfer of human chromosomes. Purified A9 microcells were fused with SPM-3T3 cells, and the microcell hybrids were selected in medium containing G418 antibiotics. The microcell hybrids that contained human chromosomes 9, 11, or 18 in a majority of cells were examined. The accumulation of intracellular cholesterol in the microcell hybrids containing a chromosome 18 decreased markedly, whereas in the microcell hybrids containing either chromosomes 9 or 11 it was similar to that in SPM3T3 cells. The SPM-3T3 cells with an intact chromosome 18 were further passaged and subcloned. Clones which again accumulated intracellular cholesterol had concurrently lost the introduced chromosome 18. The abnormal accumulation was associated with a decrement in the esterification of exogenous cholesterol. These findings suggest that the gene responsible for the abnormal cholesterol metabolism in the spm/spm mice can be restored by a hu man chromosome 18. The gene was tentatively mapped on 18pter→18p11.3 or 18q21.3→qter that was lost during subcloning, thereby resulting in reaccumulation of the intracellular cholesterol.

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References

  • Blanchette-Mackie EJ, Dwyer NK, Amende LM, Kruth HS, Butler JD, Sokol J, Comly ME, Vanier MT, August JT, Brady RO, Pentchev PG (1988) Type C Niemann-Pick disease: low-density lipoprotein uptake is associated with premature cholesterol accumulation in the Golgi complex and excessive cholesterol storage in lysosomes. Proc Natl Acad Sci USA 85:8022–8026

    Google Scholar 

  • Boylan KB, Takahashi N, Diamond M, Hood LE, Prusiner SB (1987) DNA length polymorphism located 5′ to the human myelin basic protein gene. Am J Hum Genet 40:387–400

    Google Scholar 

  • Fearon ER, Cho KR, Nigro JM, Kern SE, Simons JW, Ruppert JM, Hamilton SR, Preisinger AC, Homas G, Kinzler KW, Vogelstein BV (1990) Identification of a chromosome 18q gene that is altered in colorectal cancers. Science 247:49–56

    Google Scholar 

  • Fink JK, Filling-Katz MR, Sokol J, Cogan DG, Pikus A, Sonies B, Soong B, Pentchev PG, Comly ME, Brady RO, Barton NW (1989) Clinical spectrum of Niemann-Pick disease type C. Neurology 39:1040–1049

    Google Scholar 

  • Goss SJ, Harris H (1975) New method for mapping genes in human chromosomes. Nature 255:680–684

    CAS  PubMed  Google Scholar 

  • Hosoya M, Kimura C, Ogi K, Ohkubo S, Miyamoto Y, Kugoh H, Shimizu M, Onda H, Oshimura M, Arimura A, Fujino M (1992) Structure of the human pituitary adenylate cyclase activating polypeptide (PACAP) gene. Biochim Biophys Acta 1129:199–206

    Google Scholar 

  • Ishizaki K, Oshimura M, Sasaki MS, Nakamura Y, Ikenaga M (1990) Human chromosome 9 can complement UV sensitivity of xeroderma pigmentosum group A cells. Mutat Res 35:209–215

    Google Scholar 

  • Koi M, Morita H, Shimizu M, Oshimura M (1989a) Construction of mouse A9 clones containing a single human chromosome (X/Autosome translocation) via micro-cell fusion. Jpn J Cancer Res 80:122–125

    Google Scholar 

  • Koi M, Shimizu M, Morita H, Yamada H, Oshimura M (1989b) Construction of mouse A9 clones containing a single human chromosome tagged with neomycin-resistance gene via microcell fusion. Jpn J Cancer Res 80:413–418

    Google Scholar 

  • Komatsu K, Kodama S, Okumura Y, Koi M, Oshimura M (1990) Restoration of radiation resistance in ataxia telangiectasia cells by the introduction of normal human chromosome 11. Mutat Res 235:59–63

    Google Scholar 

  • Kruth HS, Vaughan M (1980) Quantification of low density lipoprotein binding and cholesterol accumulation by single human fibroblasts using fluorescence microscopy. J Lipid Res 21:123–130

    Google Scholar 

  • Kugoh MH, Koi M, Shimizu M, Yamada H, Oshimura M (1989) Technical notes for the transfer of chromosome tagged with a dominant selectable gene. Chromosome Information Service 46:25–27

    Google Scholar 

  • McKusick VA (ed) (1990) The Oxford Gird (Homology of synteny in mouse and man). In: Mendelian inheritance in man, 9th edn. John Hopkins University Press, Baltimore London, p 167

  • Miyawaki S, Mitsuoka S, Sakiyama T, Kitagawa T (1982) Sphingomyelinosis, a new mutation in the mouse. J Hered 73:257–263

    Google Scholar 

  • O'Connell P, Lathrop GM, Leppert M, Nakamura Y, Muller U, Lalouel J-M, White R (1988) Twelve loci form a continuous linkage map for human chromosome 18. Genomics 3:367–372

    Google Scholar 

  • Ohno K, Nanba E, Miyawaki S, Sakiyama T, Kitagawa T, Takeshita K (1992) A cell line derived from sphingomyelinosis mouse shows alterations in intracellular cholesterol metabolism similar to those in type C Niemann-Pick disease. Cell Struct Funct 17:229–235

    Google Scholar 

  • Oshimura M, Gilmer TM, Barrett JC (1985) Nonrandom loss of chromosome 15 in Syrian hamster tumors induced by v-Ha-ras plus v-myc oncogenes. Nature 316:636–639

    Google Scholar 

  • Oshimura M, Kugoh HM, Shimizu M, Yamada H, Hashiba H, Horikawa I, Sasaki M (1990) Multiple chromosomes carrying tumor suppressor activity, via microcell-mediated chromosome transfer, for various tumor cell lines. In: Knudson AG, Stanbridge EJ, Sugimura T, Terada M, Watanabe S (eds) Genetic basis for carcinogenesis: tumor suppressor genes and oncogenes. Japan Science Society Press, Tokyo/Taylor & Francis, London, pp 249–257

    Google Scholar 

  • Pentchev PG, Comly ME, Kruth HS, Vanier MT, Wenger DA, Patel S, Brady RO (1985) A defect in cholesterol esterification in Niemann-Pick disease (type C) patients. Proc Natl Acad Sci USA 82:8247–8251

    Google Scholar 

  • Pereira LV, Desnick RJ, Adler DA, Disteche CM, Schuchman EH (1991) Regional assignment of the human acid sphingomyelinase gene (SMPD1) by PCR analysis of somatic cell hybrids and in situ hybridization to 11p15.1→p15.4. Genomics 9:229–234

    Google Scholar 

  • Reed KC, Mann DA (1985) Rapid transfer of DNA from agarose gels to nylon membranes. Nucleic Acids Res 13:7207–7221

    Google Scholar 

  • Rubin CM, Houck CM, Deininger PL, Friedmann T, Schmid CW (1980) Partial nucleotide sequence of the 300-nucleotide interspersed repeated human DNA sequences. Nature 284:372–374

    Google Scholar 

  • Sakai Y, Miyawaki S, Shimizu A, Ohno K, Watanabe T (1991) A molecular genetic linkage map of mouse chromosome 18, including spm, Grl-1, Fim-2/c-fms, and Mbp. Biochem Genet 29:103–113

    Google Scholar 

  • Sakiyama T, Tsuda M, Kitagawa T, Fujita R, Miyawaki S (1982) A lysosomal storage disorder in mice: a model of Niemann-Pick disease. J Inherited Metab Dis 5:239–240

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (eds) (1989) Molecular cloning, 2nd edn. Cold Spring Harbor Laboratory Press. Cold Spring Harbor, NY

    Google Scholar 

  • Shay JW, Clark MA (1979) Nuclear control of tumorigenicity in cells reconstructed by PEG-induced fusion of cell fragments. J Supramol Struct 11:33–49

    Google Scholar 

  • Shimizu M, Yokota J, Mori N, Shuin T, Shinoda M, Terada M, Oshimura M (1990) Introduction of normal chromosome 3p modulates in the tumorigenicity of a human renal cell carcinoma cell line YCR. Oncogene 5:185–194

    Google Scholar 

  • Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517

    CAS  PubMed  Google Scholar 

  • Sokol J, Blanchette-Mackie EJ, Kruth HS, Dwyer NK, Amende LM, Butler JD, Robinson E, Patel S, Brady RO, Comly ME. Vanier MT, Pentchev PG (1988) Type C Niemann-Pick disease: lysosomal accumulation and defective intracellular morbilization of LDL-cholesterol. J Biol Chem 263:3411–3417

    Google Scholar 

  • Sparkes RS, Sasaki H, Mohandas T, Yoshioka K, Klisak I, Sasaki Y, Heinzmann C, Simon MI (1987) Assignment of the prealbumin (PALB) gene (familial amyloidotic polyneuropathy) to human chromosome region 18q11.2→q12. 1 Human Genet 75:151–154

    Google Scholar 

  • Spence MW, Callahan JW (1989) Sphingomyeline-cholesterol lipidoses: the Niemann-Pick group of disease. In: Scriver CR, Beaudet AL, Sly VS, Valle D (eds) The metabolic basis of inherited disease, 6th edn. McGraw-Hill, New York, pp 1655–1676

    Google Scholar 

  • Vanier MT, Rousson R, Zeitouni R, Pentchev PG, Louisot P (1986) Sphingomyelinase and Niemann-Pick diseae. In: Freysz L, Dreyfus H, Massarelli R, Gatt S (eds) Enzymes of lipid metabolism II. Plenum Press, New York, pp 791–802

    Google Scholar 

  • Vanier MT, Wenger DA, Comly ME, Rousson R, Brady RO, Pentchev PG (1988) Niemann-Pick disease group C: clinical variability and diagnosis based on defective cholesterol esterification. A collaborative study on 70 patients. Clin Genet 33:331–348

    Google Scholar 

  • Vanier MT, Pentchev P, Rodriguez-Lafrasse C, Rousson R (1991) Niemann-Pick disease type C: an update. J Inherited Metab Dis 14:580–595

    Google Scholar 

  • Yoshida MC, Ikeuchi T, Sasaki M (1975) Differential staining of parental chromosomes in interspecific cell hybrids with a combined quinacrine and 33258 Hoechst technique. Proc Jpn Acad 51:184–187

    Google Scholar 

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Authors and Affiliations

  1. Department of Molecular and Cell Genetics, School of Life Sciences, Tottori University, Nishimachi 86, Yonago 683, Tottori, Japan

    Akihiro Kurimasa & Mitsuo Oshimura

  2. Division of Child Neurology, Institute of Neurological Science, Faculty of Medicine, Tottori University, Yonago 683, Tottori, Japan

    Kousaku Ohno

Authors
  1. Akihiro Kurimasa
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  2. Kousaku Ohno
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  3. Mitsuo Oshimura
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Kurimasa, A., Ohno, K. & Oshimura, M. Restoration of the cholesterol metabolism in 3T3 cell lines derived from the sphingomyelinosis mouse (spm/spm) by transfer of a human chromosome 18. Hum Genet 92, 157–162 (1993). https://doi.org/10.1007/BF00219684

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  • DOI: https://doi.org/10.1007/BF00219684

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