Nucleic Acids Research, Vol 25, Issue 4 743-749, Copyright © 1997 by Oxford University Press
F Ahne, B Jha and F Eckardt-Schupp
In wild-type yeast, the repair of a 169 bp double-strand gap induced by the
restriction enzymes ApaI and NcoI in the URA3gene of the shuttle vector
YpJA18 occurs with high fidelity according to the homologous chromosomal
sequence. In contrast, only 25% of the cells of rad5-7 and rad5 Delta
mutants perform correct gap repair. As has been proven by sequencing of the
junction sites, the remaining cells recircularise the gapped plasmids by
joining of the non-compatible, non-homologous ends. Thus, regarding the
repair of DNA double-strand breaks, the rad5 mutants behave like mammalian
cells rather than budding yeast. The majority of the end joined plasmids
miss either one or both of the 3'and 5'protruding single-strands of the
restriction ends completely and have undergone blunt-end ligation
accompanied by fill-in DNA synthesis. These results imply an important role
for the Rad5 protein (Rad5p) in the protection of protruding single-strand
ends and for the avoidance of non-homologous end joining during repair of
double-strand gaps in budding yeast. Alternatively, the Rad5p may be an
accessory factor increasing the efficiency of homologous recombination in
yeast, however, the molecular mechanism of Rad5p function requires further
investigation.
ARTICLES
The RAD5 gene product is involved in the avoidance of non-homologous end-joining of DNA double strand breaks in the yeast Saccharomyces cerevisiae
Institut fur Strahlenbiologie, GSF-Forschungszentrum fur Umwelt und Gesundheit GmbH, Neuherberg, Germany.
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