Document Detail

Amplified KpnL repetitive DNA sequences in homogeneously staining regions of a human melanoma cell line.
MedLine Citation:
PMID:  6200639     Owner:  NLM     Status:  MEDLINE    
The human melanoma cell line MeWo was found to contain two populations of cells, one containing 83 chromosomes (hypotetraploid) and the other containing 43 chromosomes (hypodiploid). All of the hypodiploid cells, but none of the hypotetraploid cells, contained chromosomes with long homogeneously staining regions (HSR's) when examined with quinacrine fluorescence. These long HSR's on an X-chromosome and derivative chromosome 15, produced characteristic patterns of positive- and negative-staining areas along the HSR's with both conventional Giemsa (G) staining and C-banding. The C- and G-positive regions stained with distamycin A-DAPI, which is specific for the centromeric heterochromatin of chromosomes 1, 9, 15p, 16, and Y. DNA extracted from MeWo cells and digested with the restriction enzymes KpnL or Sau3A exhibited marked amplification of a 1.8-kilobase fragment. The amplified Sau3A fragment (D15Z1) was cloned, mapped, and partially sequenced. The sequenced region contained a five-base-pair repeat unit (adenine-adenine-thymine-guanine-guanine) that has undergone considerable divergence. Estimates of the size of the HSR's and the amount of amplified DNA suggested that each HSR contained at least 30,000 copies of the 1.8-kb KpnL fragment, representing about 50% of each HSR. The amplified sequence was identified as one member of the previously described KpnL family of repeated sequences. In situ hybridization of D15Z1 to MeWo metaphase chromosomes resulted in heavy labeling over both HSR's. These data suggested that centromeric heterochromatin and neighboring sequences on chromosome 15 have been amplified and some of this material translocated to the X-chromosome.
M C Simmons; J Maxwell; T Haliotis; M J Higgins; J C Roder; B N White; J J Holden
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of the National Cancer Institute     Volume:  72     ISSN:  0027-8874     ISO Abbreviation:  J. Natl. Cancer Inst.     Publication Date:  1984 Apr 
Date Detail:
Created Date:  1984-05-14     Completed Date:  1984-05-14     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7503089     Medline TA:  J Natl Cancer Inst     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  801-8     Citation Subset:  IM    
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MeSH Terms
DNA Restriction Enzymes / metabolism
DNA, Neoplasm / isolation & purification,  metabolism
Gene Amplification
Melanoma / genetics*,  metabolism
Nucleic Acid Hybridization
Placenta / ultrastructure
Repetitive Sequences, Nucleic Acid*
Staining and Labeling
Translocation, Genetic
Reg. No./Substance:
0/DNA, Neoplasm; EC 3.1.21.-/DNA Restriction Enzymes

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