Isolation and structural analysis of a 1.2-megabase N-myc amplicon from a human neuroblastoma

Sandra S. Schneider, Jill Reiter, Barbara A. Zehnbauer, Patricia Taillon-Miller, Denis L. Le Paslier, Bert Vogelstein, Garrett M. Brodeur

Research output: Contribution to journalArticle

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Abstract

Oncogene amplification is observed frequently in human cancers, but little is known about the mechanism of gene amplification or the structure of amplified DNA in tumor cells. We have studied the N-myc amplified domain from a representative neuroblastoma cell line, SMS-KAN, and compared the map of the amplicon in this cell line with that seen in normal DNA. The SMS-KAN cell line DNA was cloned into yeast artificial chromosomes (YACs), and clones were identified by screening the YAC library with amplified DNA probes that were obtained previously (B. Zehnbauer, D. Small, G. M. Brodeur, R. Seeger, and B. Vogelstein, Mol. Cell. Biol. 8:522-530, 1988). In addition, YAC clones corresponding to the normal N-myc locus on chromosome 2 were obtained by screening two normal human YAC libraries with these probes, and the restriction maps of the two sets of overlapping YACs were compared. Our results suggest that the amplified domain in this cell line is a ∼1.2-Mb circular molecule with a head-to-tail configuration, and the physical map of the normal N-myc locus generally is conserved in the amplicon. These results provide a physical map of the amplified domain of a neuroblastoma cell line that has de novo amplification of an oncogene. The head-to-tail organization, the general conservation of the normal physical map in the amplicon, and the extrachromosomal location of the amplified DNA are most consistent with the episome formation-plus-segregation mechanism of gene amplification in these tumors.

Original languageEnglish (US)
Pages (from-to)5563-5570
Number of pages8
JournalMolecular and Cellular Biology
Volume12
Issue number12
StatePublished - Dec 1992
Externally publishedYes

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Yeast Artificial Chromosomes
Neuroblastoma
Cell Line
Gene Amplification
DNA
Oncogenes
Human Artificial Chromosomes
Clone Cells
Neoplasms
Chromosomes, Human, Pair 2
DNA Probes
Plasmids

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Schneider, S. S., Reiter, J., Zehnbauer, B. A., Taillon-Miller, P., Le Paslier, D. L., Vogelstein, B., & Brodeur, G. M. (1992). Isolation and structural analysis of a 1.2-megabase N-myc amplicon from a human neuroblastoma. Molecular and Cellular Biology, 12(12), 5563-5570.

Isolation and structural analysis of a 1.2-megabase N-myc amplicon from a human neuroblastoma. / Schneider, Sandra S.; Reiter, Jill; Zehnbauer, Barbara A.; Taillon-Miller, Patricia; Le Paslier, Denis L.; Vogelstein, Bert; Brodeur, Garrett M.

In: Molecular and Cellular Biology, Vol. 12, No. 12, 12.1992, p. 5563-5570.

Research output: Contribution to journalArticle

Schneider, SS, Reiter, J, Zehnbauer, BA, Taillon-Miller, P, Le Paslier, DL, Vogelstein, B & Brodeur, GM 1992, 'Isolation and structural analysis of a 1.2-megabase N-myc amplicon from a human neuroblastoma', Molecular and Cellular Biology, vol. 12, no. 12, pp. 5563-5570.
Schneider SS, Reiter J, Zehnbauer BA, Taillon-Miller P, Le Paslier DL, Vogelstein B et al. Isolation and structural analysis of a 1.2-megabase N-myc amplicon from a human neuroblastoma. Molecular and Cellular Biology. 1992 Dec;12(12):5563-5570.
Schneider, Sandra S. ; Reiter, Jill ; Zehnbauer, Barbara A. ; Taillon-Miller, Patricia ; Le Paslier, Denis L. ; Vogelstein, Bert ; Brodeur, Garrett M. / Isolation and structural analysis of a 1.2-megabase N-myc amplicon from a human neuroblastoma. In: Molecular and Cellular Biology. 1992 ; Vol. 12, No. 12. pp. 5563-5570.
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