Repair and misrepair of site-specific DNA double-strand breaks by human cell extracts

Mutat Res. 1993 May;299(3-4):251-9. doi: 10.1016/0165-1218(93)90101-i.

Abstract

The rejoining by human cell extracts of a double-strand break induced by endonuclease treatment at one of several sites within a small DNA molecule was studied. Rejoining was found at each of 8 sites tested, but the rejoin efficiency varied with the nature of the break (e.g., breaks with cohesive ends were rejoined more efficiently than blunt-ended breaks). Extracts from primary and immortalized cell lines, as well as those from individuals with ataxia telangiectasia (A-T), showed the same pattern of relative rejoin efficiencies. However, mis-rejoining varied with the cell extract used, and was particularly elevated with two immortalized A-T cell lines. Mixing experiments showed that the mis-rejoining property of extracts could act in a semi-dominant fashion, depending on the individual efficiencies of the component extracts. The mis-rejoin mechanism involved deletion at sites of short direct repeats at various distances from the initial break site. A model of deletion formation (the strand-exposure and repair model) is restated to explain the sequence repeat dependence found, and is compared to models of homologous DNA recombination.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ataxia Telangiectasia
  • Base Composition
  • Base Sequence
  • Cell Line
  • Cell-Free System
  • Cloning, Molecular / methods
  • DNA / genetics*
  • DNA Damage*
  • DNA Repair*
  • DNA Restriction Enzymes / metabolism
  • Humans
  • Models, Genetic
  • Molecular Sequence Data
  • Mutagenesis
  • Plasmids
  • Restriction Mapping
  • Substrate Specificity

Substances

  • DNA
  • DNA Restriction Enzymes