Nucleic Acids Research, 1992, Vol. 20, No. 23 6183-6188
© 1992
MOLECULAR BIOLOGY |
A mechanism for deletion formation in DNA by human cell extracts: the involvement of short sequence repeats
MRC Radiobiology Unit Chilton, Didcot, Oxon OX11 0RD, UK
Received October 13, 1992. Revised November 5, 1992. Accepted November 5, 1992.
DNA molecules carrying a site-specific double-strand break were exposed to nuclear extracts from human cell lines. It was shown previously that breaks could be rejoined correctly by human extracts, but that a proportion of the rejoined molecules had suffered deletions and insertions. The ‘mis-rejoined’ proportion was higher with cell extracts from an individual with the disorder ataxia-telanglectasia than with normal cell extracts. We now show by sequence analysis that deletions in extract-treated molecules occur exclusively between short direct repeats (2–6 base pairs). A misrejoined molecule containing an insertion of 300 bp also had a repeat-based deletion at the same site. A number of different direct repeats are involved; however, some clustering of these occurs especially on the upstream side of the initial breakpoint. These data are most simply interpreted in terms of a model of deletion formation involving single-strand exposure and repair, perhaps with the action of other DNA-metabolising enzymes influencing the frequency with which some repeats are involved.
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