Abstract
A cultured line of neuroblastoma cells (NB) was found to contain double minute chromosomes (DMs). DMs have been reported to be acentric and, therefore, to be segregated randomly into daughter cells without separating their sister elements. When NB cells were fused with Chinese hamster metaphase cells, prematurely condensed chromosomes (PCCs) were induced. DMs seen together with G2 PCCs appeared to be closely paired, dot-like structures resembling DMs observable in metaphase cells. In contrast, DMs in G1 cells showed a tendency to become single as the stage progressed so that the majority of DMs in late G1 cells were actually no longer double. DMs in S-phase cells, however, again appeared double. These results clearly indicate why DMs are invariably double and never assume a quadruple configuration in metaphase cells in spite of their non-disjunctional segregation at anaphase. Such a characteristic mode of DM replication was further confirmed by a 5-bromo-2′-deoxyuridine (BrdUrd) labeling experiment: when NB cells were exposed to BrdUrd for two successive rounds of DNA replication prior to PCC induction, half of the resulting single G1 minutes as well as G1 PCCs stained dark and the other half stained light after staining for sister chromatid differentiation.
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References
Alitalo K, Schwab M, Lin CC, Varmus HE, Bishop JM (1983) Homogeneously staining chromosomal regions contain amplified copies of an abundantly expressed cellular oncogene (c-myc) in malignant neuroendocrine cells from a human colon carcinoma. Proc Natl Acad Sci USA 80:1707–1711
Bahr G, Gilbert F, Balaban G, Engler W (1983) Homogeneously staining regions and double minutes in a human cell line: chromatin organization and DNA content. J Natl Cancer Inst 71:657–661
Barker PE (1982) Double minutes in human tumor cells. Cancer Genet Cytogenet 5:81–94
Barker PE, Hsu TC (1978) Are double minute chromosomes? Exp Cell Res 113:456–458
Barker PE, Hsu TC (1979) Double minutes in human carcinoma cell lines, with special reference to breast tumors. J Natl Cancer Inst 62:257–262
Barker PE, Stubblefield E (1979) Ultrastructure of double minute from a human tumor cell line. J Cell Biol 83:663–666
Barker PE, Drwinga HL, Hittelman WN, Maddox A-M (1980) Double minutes replicate once during S phase of the cell cycle. Exp Cell Res 130:353–360
Biedler JL, Spengler BA (1976) Metaphase chromosome anomaly: Association with drug resistance and cell-specific products. Science 191:185–187
Biedler JL, Meyers MB, Spengler BA (1983) Homogeneously staining regions and double minute chromosomes, prevalent cytogenetic abnormalities of human neuroblastoma cells. Adv Cell Neurobiol 4:267–307
Brown PC, Beverley SM, Schimke RT (1981) Relationship of amplified dihydrofolate reductase genes to double minute chromosomes in unstably resistant mouse fibroblast cell lines. Mol Cell Biol 1:1077–1083
Collins SJ, Groudine MT (1983) Rearrangement and amplification of c-abl sequences in human chronic myelogenous leukemia cell line K-562. Proc Natl Acad Sci USA 80:4813–4817
Cowell JK (1982) Double minutes and homogeneously staining regions: Gene amplification in mammalian cells. Annu Rev Genet 16:21–16
Dalla-Favera R, Wong-Staal F, Gallo RC (1982) Oncogene amplification in promyelocytic leukaemia cell line HL-60 and primary leukaemic cells of the same patient. Nature 299:61–63
Emanuel BS, Balaban G, Boid JP, Grossman A, Negishi M, Parmiter A, Glick MC (1985) N-myc amplification in multiple homogeneously staining regions in two human neuroblastomas. Proc Natl Acad Sci USA 82:3736–3740
Goyanes VJ, Mendez J (1982) Extracentromeric connections between sister chromatids demonstrated in human chromosomes induced to condense asymmetrically. Hum Genet 62:324–326
Hamkalo BA, Farnham PJ, Hohnston R, Schimke RT (1985) Ultrastructural features of minute chromosomes in a methotrexate resistant mouse 3T3 cell line. Proc Natl Acad Sci USA 82:1126–1130
Hamlin JL, Milbrandt JD, Heintz NH, Azizkhan JC (1984) DNA sequence amplification in mammalian cells. Int Rev Cytol 90:31–77
Hanks SK, Brown DB, Rao PN (1982) Induction of premature chromosome condensation at high frequency following polyethylene glycol-mediated fusion of lectin-bound cells. Exp Cell Res 138:215–219
Jack EM, Waters JJ, Harrison CJ (1987) A scanning electron microscopy study of double minutes from a human tumor cell line. Cytogenet Cell Genet 44:49–52
Kanda N, Schreck R, Alt F, Bruns G, Baltimore D, Latt S (1983) Isolation of amplified DNA sequences from IMR-32 human neuroblastoma cells: facilitation by fluorescence-activated flow sorting of metaphase chromosomes. Proc Natl Acad Sci USA 80:4069–4073
Kaneko Y, Kanda N, Maseki N, Sakurai M, Tsuchida Y, Takeda T, Okabe I, Sakurai M (1987) Different karyotypic patterns in early and advanced stage neuroblastomas. Cancer Res 47:311–318
Kohl NE, Kanda N, Schreck RR, Bruns G, Latt SA, Gilbert F, Alt FW (1983) Transposition and amplification of oncogenerelated sequences in human neuroblastomas. Cell 35:359–367
Levan A, Levan G (1978) Have double minutes functioning centromeres? Hereditas 88:81–92
Levan A, Levan G, Mandahl N (1978) A new chromosome type replacing the double minutes in a mouse tumor. Cytogenet Cell Genet 20:12–23
Levan G, Mandahl N, Bregula U, Klein G, Levan A (1976) Double minute chromosomes are not centromeric regions of the host chromosomes. Hereditas 83:83–90
Lica LM, Narayanswami S, Hamkalo BA (1986) Mouse satellite DNA, centromere structure, and sister chromatid pairing. J Cell Biol 103:1145–1151
Montgomery KT, Biedler JL, Spengler BA, Melera PW (1983) Specific DNA sequence amplification in human neuroblastoma cells. Proc Natl Acad Sci USA 80:5724–5728
Nunberg JH, Kaufman RJ, Schimke RT, Urlarub G, Chasin LA (1978) Amplified dihydrofolate reductase genes are localized to a homogeneously staining region of single chromosome in a methotrexate resistant Chines hamster ovary cell line. Proc Natl Acad Sci USA 75:5553–5556
Rattner JB, Lin CC (1984) Ultrastructural organization of double minute chromosomes and HSR regions in human colon carcinoma cells. Cytogenet Cell Genet 38:176–181
Schimke RT (1982) Studies on gene duplications and amplifications — An historical perspective. In: Schimke RT (ed) Gene amplification. Cold Spring Harbor Laboratory, NY, pp 1–6
Schwab M, Alitalo K, Varmus HE, Bishop JM, George D (1983) A cellular oncogene (c-ki-ras) is amplified, overexpressed, and located with karyotypic abnormalities in mouse adrenocortical tumor cells. Nature 303:497–501
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Takayama, S., Uwaike, Y. Analysis of the replication mode of double minutes using the PCC technique combined with BrdUrd labeling. Chromosoma 97, 198–203 (1988). https://doi.org/10.1007/BF00292961
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DOI: https://doi.org/10.1007/BF00292961