Abstract
An H4-IIE-C3 hepatoma cell line derived from an ACI rat has been shown to have differentially stained regions attached to the short arms of chromosomes 3, 11 and 13 and the long arm of an unidentified small chromosome. There is cell to cell variability in the number and size of the differentially stained regions, which contain, on the average, about 5% of the total DNA. A series of secondary constrictions occur at intervals along the length of each differentially stained region. These stain with silver by the Ag-AS method, indicating that the differentially stained regions contain sites of active 45S ribosomal precursor RNA transcription. In situ hybridization to metaphase chromosomes shows that the hepatoma cells have a 10 fold increase in DNA coding for 18S and 28S ribosomal RNA, 90% of it located in the differentially stained regions, and no change in the number of genes coding for 5S RNA. These results have been confirmed by filter disc hybridization.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Alt, F.W., Kellems, R.E., Bertino, J.R., Schimke, R.T.: Selective multiplication of dihydrofolate reductase genes in methotrexate-resistant variants of cultured murine cells. J. biol. Chem. 253, 1357–1370 (1978)
Balaban-Malenbaum, G., Gilbert, F.: Double minute chromosomes and the homogeneously staining regions in chromosomes of a human neuroblastoma cell line. Science 198, 739–740 (1977)
Balazs, I., Szabo, P., Siniscalco, M.: Somatic Cell Genet. (in press, 1978)
Biedler, J.L., Spengler, B.A.: A novel chromosome abnormality in human neuroblastoma and antifolate-resistant Chinese hamster cell lines in culture. J. nat. Cancer Inst. 57, 683–695 (1976)
Bird, A.P., Southern, E.M.: Use of restriction enzymes to study eukaryotic DNA methylation: I. The methylation pattern in ribosomal DNA from Xenopus laevis. J. molec Biol. 118, 27–47 (1978)
Birnstiel, M.L., Sells, B.H., Purdom, I.F.: Kinetic complexity of RNA molecules. J. molec Biol. 63, 21–39 (1972)
Bobrow, M., Collacott, H.E., Madan, K.: Chromosome banding with acridine orange. Lancet 1972ii, 1311
Brown, D.D., Dawid, I.B.: Specific gene amplification in oocytes. Science 160, 272–280 (1968)
Chesterton, C.J., Humphrey, S.M., Butterworth, P.H.W.: Comparison of the multiple deoxyribonucleic acid-dependent ribonucleic acid polymerase forms of whole rat liver and a minimal-deviation rat hepatoma cell line. Biochem. J. 126, 675–681 (1972)
Committee for a standardized karyotype of Rattus norvegicus: Standard karyotype of the Norway rat, Rattus norvegicus. Cytogenet. Cell Genet. 12, 199–205 (1973)
Cory, S., Adams, J.M.: A very large repeating unit of mouse DNA containing the 18S, 28S and 5.8S rRNA genes. Cell 11, 795–805 (1977)
Goodpasture, C., Bloom, S.E.: Visualization of nucleolar organizer regions in mammalian chromosomes using silver staining. Chromosoma (Berl.) 53, 37–50 (1975)
Hakala, M.T., Zakrzewski, S.F., Nichol, C.A.: Relation of folic acid reduction to amethopterin resistance in cultured mammalian cells. J. biol. Chem. 236, 952–958 (1961)
Handbook of Biochemistry, 2nd edition (H.A. Saber, ed.). Sect. H, pp. 112–115. Cleveland: CRC Press 1970
Honjo, T., Reeder, R.H.: Preferential transcription of Xenopus laevis ribosomal RNA in interspecies hybrids between Xenopus laevis and Xenopus mulleri. J. molec. Biol. 80, 217–228 (1973)
Lee, M.R.: A widela applicable technic for direct processing of bone marrow for chromosomes of vertebrates. Stain Technol. 44, 155–158 (1969)
Levan, A., Levan, G., Mandahl, N.: A new chromosome type replacing the double minutes in a mouse tumor. Cytogenet. Cell Genet. 20, 12–23 (1978)
Levan, G., Levan, A.: Specific chromosome changes in malignancy: studies in rat sarcomas induced by two polycyclic hydrocarbons. Hereditas (Lund) 79, 161–198 (1975)
Matsushima, T., Yosida, T.H.: Change of stemline karyotypes in Yosida sarcoma by appearance of peculiar marker chromosomes. Gann 62, 389–394 (1971)
Miller, D.A., Breg, W.R., Warburton, D., Dev, V.G., Miller, O.J.: Regulation of rRNA gene expression in a human familial 14 p+marker chromosome. Hum. Genet. 43, 289–297 (1978)
Miller, D.A., Dev, V.G., Borek, C., Miller, O.J.: The quinacrine fluorescent and Giemsa banded karyotype of the rat, Rattus norvegicus, and banded chromosome analysis of transformed and malignant rat liver cell lines. Cancer Res. 32, 2375–2382 (1972)
Miller, D.A., Dev, V.G., Tantravahi, R., Miller, O.J.: Suppression of human nucleolus organizer activity in mouse-human somatic hybrid cells. Exp. Cell Res. 101, 235–243 (1976a)
Miller, D.A., Tantravahi, R., Dev, V.G., Miller, O.J.: Q- and C-band chromosome variants in inbred strains of Mus musculus. Genetics 84, 67–75 (1976b)
Miller, O.J., Miller, D.A., Dev, V.G., Tantravahi, R., Croce, C.M.: Expression of human and suppression of mouse nucleolus organizer activity in mouse-human somatic cell hybrids. Proc. nat. Acad. Sci. (Wash.) 73, 4531–4535 (1976 c)
Miller, O.J., Miller, D.A., Kouri, R.E., Allerdice, P.W., Dev, V.G., Grewal, M.S., Hutton, J.J.: Identification of the mouse karyotype by quinacrine fluorescence and tentative assignment of seven linkage groups. Proc. nat. Acad. Sci. (Wash.) 68, 1530–1533 (1971)
Mullen, V.T., Barnett, C.A.: Banded karyotypes of H-4-IIE-C3 rat hepatoma cells grown in vitro. In Vitro 12, 658–664 (1976)
Nielsen, K.: Chromosome evolution in the Ehrlich-Lettré complex of hyperdiploid mouse ascites tumors: Results from seven laboratory strains. Hereditas (Lund) 84, 77–108 (1976)
Oshimura, M., Sasaki, M., Makino, S.: Chromosomal banding patterns in primary and transplanted veneral tumors of the dog. J. nat. Cancer Inst. 51, 1197–1203 (1973)
Perry, R.P., Kelley, D.E.: Persistent synthesis of 5S RNA when production of 28S and 18S ribosomal RNA is inhibited by low doses of Actinomycin D. J. cell. Physiol. 72, 235–246 (1968)
Pitot, H.C., Peraino, C., Morse, P.A., Jr., Potter, V.R.: Hepatomas in tissue culture compared with adpating liver in vivo. Nat. Cancer Inst. Monogr. 13, 229–242 (1964)
Procunier, J.D., Tartof, K.D.: Genetic analysis of the 5S RNA genes in Drosophila melanogaster. Genetics 81, 515–523 (1975)
Ritossa, F.M.: Unstable redundancy of genes for ribosomal RNA. Proc. nat. Acad. Sci. (Wash.) 60, 509–516 (1968)
Sasaki, M., Oshimura, M., Makino, S., Koike, T., Itoh, M., Watanabe, F., Tanaka, N.: Further karyological evidence for contagiousness and common origin of canine venereal tumors. Proc. Jap. Acad. 50, 636–640 (1974)
Schwartz, L.B., Roeder, R.G.: Purification and subunit structure of deoxyribonucleic acid-dependent ribonucleic acid polymerase I from mouse melanoma MOPC 315. J. biol. Chem. 249, 5898–5906 (1974)
Sehested, J.: A simple method for R banding of human chromosomes, showing a pH-dependent connection between R and G bands. Humangenetik 21, 55–58 (1974)
Sun, N.C., Chu, E.H.Y., Chang, C.C.: Staining method for the banding patterns of human mitotic chromosomes. Caryologia (Firenze) 27, 315–324 (1974)
Szabo, P., Lee, M.R., Elder, F.B., Prensky, W.: Localization of 5S RNA and rRNA genes in the Norway rat. Chromosoma (Berl.) 65, 161–172 (1978)
Tantravahi, R., Miller, D.A., D'Ancona, G., Croce, C.M., Miller O.J.: Location of rRNA genes in three inbred strains of rat and suppression of rat rRNA activity in rat-human somatic cell hybrids. Exp. Cell Res. (in press, 1979)
Weinmann, R.: Regulation of ribosomal RNA and 5S RNA synthesis in Drosophila melanogaster: I. Bobbed mutants. Genetics 72, 267–276 (1972)
Yu, L.C., Szabo, P., Borun, T.W., Prensky, W.: The localization of the genes coding for histone H4 in human chromosomes. Cold Spr. Harb. Symp. quant. Biol. 42, 1101–1105 (1978)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Miller, O.J., Tantravahi, R., Miller, D.A. et al. Marked increase in ribosomal RNA gene multiplicity in a rat hepatoma cell line. Chromosoma 71, 183–195 (1979). https://doi.org/10.1007/BF00292822
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00292822