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PMID: 8423817 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Acquisition of telomere repeat sequences by transfected DNA integrated at the site of a chromosome break.

Molecular and cellular biology ·Vol. 13 ·No. 2 ·1993-02-00 ·Pages 977-83

Murnane JP, Yu LC

Abstract

Previous analysis of plasmid DNA transfected into 108 cell clones demonstrated extensive polymorphism near the integration site in one clone. This polymorphism was apparent by Southern blot analysis as diffuse bands that extended over 30 kb. In the present study, nucleotide sequence analysis of cloned DNA from the integration site revealed telomere repeat sequences at the ends of the integrated plasmid DNA. The telomere repeat sequences at one end were located at the junction between the plasmid and cell DNA. The telomere repeat sequences at the other end were located in the opposite orientation in the polymorphic region and were shown by digestion with BAL 31 to be at the end of the chromosome. Telomere repeat sequences were not found at this location in the plasmid or parent cell DNA. Although the repeat sequences may have been acquired by recombination, a more likely explanation is that they were added to the ends of the plasmid by telomerase before integration. Comparison of the cell DNA before and after integration revealed that a chromosome break had occurred at the integration site, which was shown by fluorescent in situ hybridization to be located near the telomere of chromosome 13. These results demonstrate that chromosome breakage and rearrangement can result in interstitial telomere repeat sequences within the human genome. These sequences could promote genomic instability, because short repeat sequences can be recombinational hotspots. The results also show that DNA rearrangements involving telomere repeat sequences can be associated with chromosome breaks. The introduction of telomere repeat sequences at spontaneous or ionizing radiation-induced DNA strand breaks may therefore also be a mechanism of chromosome fragmentation.

MeSH Terms
Animals Base Sequence Blotting, Southern Cell Line Chromosomes Cloning, Molecular DNA Humans Mice Molecular Sequence Data Plasmids Repetitive Sequences, Nucleic Acid Restriction Mapping Telomere Transfection
Chemicals
DNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Murnane J P
Laboratory of Radiobiology and Environmental Health, University of California, San Francisco 94143-0750.
Yu L C
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.13.2.977-983.1993
Published
1993-02-00
Pages
977-83
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC358982
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.13.2.977-983.1993 PMC Full Text View journal details
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